Chromenone derivatives

ABSTRACT

The invention concerns chromenone derivatives of Formula I 
                         
or a pharmaceutically-acceptable salts thereof, wherein each of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , n and R 9  has any of the meanings defined hereinbefore in the description; processes for their preparation, pharmaceutical compositions containing them and their use in the manufacture of a medicament for use in the treatment of cell proliferative disorders.

This application is a continuation of U.S. patent application Ser. No.14/163,105, filed Jan. 24, 2014 (now U.S. Pat. No. 9,029,374, issued May12, 2015), which is a continuation of U.S. patent application Ser. No.13/765,850, filed Feb. 13, 2013 (now U.S. Pat. No. 8,673,906,issued Mar.18, 2014), which is a continuation of U.S. patent application Ser. No.12/909,968, filed Oct. 22, 2010 (now U.S. Pat. No. 8,399,460, issuedMar. 19, 2013), which claims the benefit under 35 U.S.C. pre-AIA §119(a)and 35 U.S.C. §119(b)-(d) of European Application No. 09306017.6, filedon Oct. 27, 2009. This application is also related to U.S. patentapplication Ser. No. 13/616,473, filed Sep. 14, 2012 and U.S. patentapplication Ser. No. 14/681,244, filed Apr. 8, 2015. Each of theforegoing applications is incorporated by reference in its entirety.

The invention concerns certain novel chromenone derivatives, orpharmaceutically-acceptable salts thereof, which possess anti-canceractivity and are accordingly useful in methods of treatment of the humanor animal body. The invention also concerns processes for themanufacture of said chromenone derivatives, pharmaceutical compositionscontaining them and their use in therapeutic methods, for example in themanufacture of medicaments for use in the prevention or treatment ofcancers in a warm-blooded animal such as man, including use in theprevention or treatment of cancer.

The present invention also relates to chromenone derivatives that areselective inhibitors of phosphoinositide (PI) 3-kinase β, and are, forexample, useful for anti-tumour therapy. Further, the present inventionalso relates to the use of chromenone derivatives of the invention thatare selective inhibitors of phosphoinositide (PI) 3-kinase β, inanti-tumour therapy. Inhibitors of PI 3-kinase β may be effective in thetreatment of tumours which are deficient in the gene PTEN (phosphataseand tensin homologue deleted on chromosome 10) and this relates to afurther feature of the invention.

In the area of cancer it has in recent years been discovered that a cellmay become cancerous by virtue of the transformation of a portion of itsDNA into an oncogene, that is a gene which, on activation, leads to theformation of malignant tumour cells (Bradshaw, Mutagenesis, 1986, 1,91). Several such oncogenes give rise to the production of peptides,which are receptors for growth factors. Activation of the growth factorreceptor complex subsequently leads to an increase in cellproliferation. It is known, for example, that several oncogenes encodetyrosine kinase enzymes and that certain growth factor receptors arealso tyrosine kinase enzymes (Yarden et al., Ann. Rev. Biochem., 1988,57, 443; Larsen et al., Ann. Reports in Med. Chem., 1989, Chpt. 13). Thefirst group of tyrosine kinases to be identified arose from such viraloncogenes, for example pp60^(v-Src) tyrosine kinase (otherwise known asv-Src), and the corresponding tyrosine kinases in normal cells, forexample pp60^(c-Src) tyrosine kinase (otherwise known as c-Src).

Receptor tyrosine kinases are important in the transmission ofbiochemical signals which initiate cell replication. They are largeenzymes which span the cell membrane and possess an extracellularbinding domain for growth factors such as epidermal growth factor (EGF)and an intracellular portion which functions as a kinase tophosphorylate tyrosine amino acids in proteins and hence to influencecell proliferation. Various classes of receptor tyrosine kinases areknown (Wilks, Advances in Cancer Research, 1993, 60, 43-73) based onfamilies of growth factors, which bind to different receptor tyrosinekinases. The classification includes Class I receptor tyrosine kinasescomprising the EGF family of receptor tyrosine kinases such as the EGF,TGFα, Neu and erbB receptors.

It is also known that certain tyrosine kinases belong to the class ofnon-receptor tyrosine kinases which are located intracellularly and areinvolved in the transmission of biochemical signals such as those thatinfluence tumour cell motility, dissemination and invasiveness andsubsequently metastatic tumour growth. Various classes of non-receptortyrosine kinases are known including the Src family such as the Src,Lyn, Fyn and Yes tyrosine kinases.

Further, it is also known that certain kinases belong to the class ofserine/threonine kinases which are located intracellularly anddownstream of tyrosine kinase activation and are involved in thetransmission of biochemical signals such as those that influence tumourcell growth. Such serine/threonine signalling pathways include theRaf-MEK-ERK cascade and those downstream of PI 3-KINASE such as PDK-1,AKT and mTOR (Blume-Jensen and Hunter, Nature, 2001, 411, 355).

It is also known that certain other kinases belong to the class of lipidkinases, which are located intracellularly and are also involved in thetransmission of biochemical signals such as those that influence tumourcell growth and invasiveness. Various classes of lipid kinases are knownincluding the aforementioned PI 3-kinase family, which is alternativelyknown as the phosphatidylinositol-3-kinase family.

It is now well understood that deregulation of oncogenes andtumour-suppressor genes contributes to the formation of malignanttumours, for example by way of increased cell proliferation or increasedcell survival. It is also now known that signalling pathways mediated bythe PI 3-kinase family have a central role in a number of cell processesincluding proliferation and survival, and deregulation of these pathwaysis a causative factor a wide spectrum of human cancers and otherdiseases (Katso et al., Annual Rev. Cell Dev. Biol., 2001, 17: 615-617and Foster et al., J. Cell Science, 2003, 116: 3037-3040).

The PI 3-kinase family of lipid kinases is a group of enzymes thatphosphorylate the 3-position of the inositol ring ofphosphatidylinositol (PI). Three major groups of PI 3-kinase enzymes areknown which are classified according to their physiological substratespecificity (Vanhaesebroeck et al., Trends in Biol. Sci., 1997, 22,267). Class III PI 3-kinase enzymes phosphorylate PI alone. In contrast,Class II PI 3-kinase enzymes phosphorylate both PI and PI 4-phosphate[abbreviated hereinafter to PI(4)P]. Class I PI 3-kinase enzymesphosphorylate PI, PI(4)P and PI 4,5-bisphosphate [abbreviatedhereinafter to PI(4,5)P2], although only PI(4,5)P2 is believed to be thephysiological cellular substrate. Phosphorylation of PI(4,5)P2 producesthe lipid second messenger PI 3,4,5-triphosphate [abbreviatedhereinafter to PI(3,4,5)P3]. More distantly related members of thissuperfamily are Class IV kinases such as mTOR and DNA-dependent kinasethat phosphorylate serine/threonine residues within protein substrates.The most studied and understood of these lipid kinases are the Class IPI 3-kinase enzymes.

Class I PI 3-kinase is a heterodimer consisting of a p110 catalyticsubunit and a regulatory subunit, and the family is further divided intoClass Ia and Class Ib enzymes on the basis of regulatory partners andmechanism of regulation. Class Ia enzymes, include PI 3-kinase β, andconsist of three distinct catalytic subunits (p110α, p110β and p110δ)that dimerise with five distinct regulatory subunits (p85α, p55α, p50α,p85β and p55γ), with all catalytic subunits being able to interact withall regulatory subunits to form a variety of heterodimers. Class Ia PI3-kinase enzymes are generally activated in response to growthfactor-stimulation of receptor tyrosine kinases, via interaction of theregulatory subunit SH2 domains with specific phospho-tyrosine residuesof the activated receptor or adaptor proteins such as IRS-1. Both p110αand p110β are constitutively expressed in all cell types, whereas p110δexpression is more restricted to leukocyte populations and someepithelial cells. In contrast, the single Class Ib enzyme consists of ap110γ catalytic subunit that interacts with a p101 regulatory subunit.Furthermore, the Class Ib enzymes are activated in response to G-proteincoupled receptor (GPCR) systems as well as by the mechanisms describedabove.

There is now considerable evidence indicating that Class Ia PI 3-kinaseenzymes, which include PI 3-kinase β, contribute to tumourigenesis in awide variety of human cancers, either directly or indirectly (Vivancoand Sawyers, Nature Reviews Cancer, 2002, 2, 489-501). For example, thep110α subunit is amplified in some tumours such as those of the ovary(Shayesteh et al., Nature Genetics, 1999, 21: 99-102) and cervix (Ma etal., Oncogene, 2000, 19: 2739-2744). Activating mutations within thecatalytic site of p110α have been associated with various other tumourssuch as those of the colorectal region and of the breast and lung(Samuels et al., Science, 2004, 304, 554). Tumour-related mutations inp85α have also been identified in cancers such as those of the ovary andcolon (Philp et al., Cancer Research, 2001, 61, 7426-7429). In additionto direct effects, it is believed that activation of Class Ia PI3-kinase contributes to tumourigenic events that occur upstream insignalling pathways, for example by way of ligand-dependent orligand-independent activation of receptor tyrosine kinases, GPCR systemsor integrins (Vara et al., Cancer Treatment Reviews, 2004, 30, 193-204).Examples of such upstream signalling pathways include over-expression ofthe receptor tyrosine kinase Erb2 in a variety of tumours leading toactivation of PI 3-kinase-mediated pathways (Harari et al., Oncogene,2000, 19, 6102-6114) and over-expression of the oncogene Ras(Kauffmann-Zeh et al., Nature, 1997, 385, 544-548). In addition, ClassIa PI 3-kinases may contribute indirectly to tumourigenesis caused byvarious downstream signalling events. For example, loss of the effect ofthe PTEN tumour-suppressor phosphatase that catalyses conversion ofPI(3,4,5)P3 back to PI(4,5)P2 is associated with a very broad range oftumours via deregulation of PI 3-kinase-mediated production ofPI(3,4,5)P3 (Simpson and Parsons, Exp. Cell Res., 2001, 264, 29-41).Furthermore, augmentation of the effects of other PI 3-kinase-mediatedsignalling events is believed to contribute to a variety of cancers, forexample by activation of Akt (Nicholson and Anderson, CellularSignalling, 2002, 14, 381-395).

In addition to a role in mediating proliferative and survival signallingin tumour cells, there is also good evidence that Class Ia PI 3-kinaseenzymes will also contribute to tumourigenesis via its function intumour-associated stromal cells. For example, PI 3-kinase signalling isknown to play an important role in mediating angiogenic events inendothelial cells in response to pro-angiogenic factors such as VEGF(Abid et al., Arterioscler. Thromb. Vasc. Biol., 2004, 24, 294-300). AsClass I PI 3-kinase enzymes are also involved in motility and migration(Sawyer, Expert Opinion Investig. Drugs, 2004, 13, 1-19), PI 3-kinaseinhibitors should provide therapeutic benefit via inhibition of tumourcell invasion and metastasis.

In addition, Class I PI 3-kinase enzymes play an important role in theregulation of immune cells with PI 3-kinase activity contributing topro-tumourigenic effects of inflammatory cells (Coussens and Werb,Nature, 2002, 420, 860-867).

These findings suggest that pharmacological inhibitors of Class I PI3-kinase enzymes should be of therapeutic value for treatment of thevarious forms of the disease of cancer comprising solid tumours such ascarcinomas and sarcomas and the leukaemias and lymphoid malignancies. Inparticular, inhibitors of Class I PI 3-kinase enzymes should be oftherapeutic value for treatment of, for example, cancer of the breast,colorectum, lung (including small cell lung cancer, non-small cell lungcancer and bronchioalveolar cancer) and prostate, and of cancer of thebile duct, bone, bladder, head and neck, kidney, liver, gastrointestinaltissue, oesophagus, ovary, pancreas, skin, testes, thyroid, uterus,cervix and vulva, and of leukaemias (including ALL and CML), multiplemyeloma and lymphomas.

Generally, investigators have explored the physiological andpathological roles of the PI 3-kinase enzyme family using theaforementioned PI 3-kinase inhibitors LY294002 and wortmannin. Althoughuse of those compounds may suggest a role for PI 3-kinase in a cellularevent, they are not sufficiently selective within the PI 3-kinase familyto allow dissection of the individual roles of the family members. Forthis reason, more potent and selective pharmaceutical PI 3-kinaseinhibitors would be useful to allow a more complete understanding of PI3-kinase function and to provide useful therapeutic agents.

In addition to tumourigenesis, there is evidence that Class I PI3-kinase enzymes play a role in other diseases (Wymann et al., Trends inPharmacological Science, 2003, 24, 366-376). Both Class Ia PI 3-kinaseenzymes and the single Class Ib enzyme have important roles in cells ofthe immune system (Koyasu, Nature Immunology, 2003, 4, 313-319) and thusthey are therapeutic targets for inflammatory and allergic indications.Inhibition of PI 3-kinase is also, as described earlier, useful to treatcardiovascular disease via anti-inflammatory effects or directly byaffecting cardiac myocytes (Prasad et al., Trends in CardiovascularMedicine, 2003, 13, 206-212). Inhibition of PI 3-kinase is also usefulto treat thrombosis. WO2004016607 provides a method of disruptingplatelet aggregation and adhesion occurring under high shear conditions,and a method for inhibiting platelet activation induced by shear, whereboth methods comprise the administration of a selective PI 3-kinase βinhibitor. WO2004016607 also provides an antithrombotic methodcomprising administering an effective amount of a selective PI 3-kinaseβ inhibitor. According to the method, specific inhibition of thrombosiscan be obtained without affecting normal haemostasis by targeting PI3-kinase β that is important for shear-induced platelet activation. Saidantithrombotic method therefore does not involve side effects caused bydisruption of normal haemostasis, such as extending of bleeding time.

Thus inhibitors of Class I PI 3-kinase enzymes, including inhibitors ofPI 3-kinase β, are expected to be of value in the prevention andtreatment of a wide variety of diseases in addition to cancer.

The compounds, i.e. the chromenone derivatives, of the invention havenow surprisingly been found to possess potent anti-tumour activity,being useful in inhibiting the uncontrolled cellular proliferation whicharises from malignant disease. Without wishing to imply that thecompounds disclosed in the present invention possess pharmacologicalactivity only by virtue of an effect on a single biological process, itis believed that the compounds provide an anti-tumour effect by way ofinhibition of Class I PI 3-kinase enzymes, particularly by way ofinhibition of the Class Ia PI 3-kinase enzymes and/or the Class Ib PI3-kinase enzyme, more particularly by way of inhibition of the Class IaPI 3-kinase enzymes, which include inhibition of PI 3-kinase β.

The compounds of the present invention are also useful in inhibiting theuncontrolled cellular proliferation which arises from variousnon-malignant diseases such as inflammatory diseases (for examplerheumatoid arthritis and inflammatory bowel disease), fibrotic diseases(for example hepatic cirrhosis and lung fibrosis), glomerulonephritis,multiple sclerosis, psoriasis, benign prostatic hypertrophy (BPH),hypersensitivity reactions of the skin, blood vessel diseases (forexample atherosclerosis and restenosis), allergic asthma,insulin-dependent diabetes, diabetic retinopathy and diabeticnephropathy.

Generally, the compounds of the present invention possess potentinhibitory activity against Class I PI 3-kinase enzymes, particularlyagainst Class Ia PI 3-kinase enzymes, including against of PI 3-kinaseβ, whilst possessing less potent inhibitory activity against tyrosinekinase enzymes such as the receptor tyrosine kinases, for example EGFreceptor tyrosine kinase and/or VEGF receptor tyrosine kinase, oragainst non-receptor tyrosine kinases such as Src. Furthermore, certaincompounds of the present invention, possess substantially better potencyagainst Class I PI 3-kinase enzymes, particularly against Class Ia PI3-kinase enzymes, including against of PI 3-kinase β, than against EGFreceptor tyrosine kinase or VEGF receptor tyrosine kinase or Srcnon-receptor tyrosine kinase. Such compounds possess sufficient potencyagainst Class I PI 3-kinase enzymes that they may be used in an amountsufficient to inhibit Class I PI 3-kinase enzymes, particularly toinhibit Class Ia PI 3-kinase enzymes, including PI 3-kinase β, whilstdemonstrating little activity against EGF receptor tyrosine kinase orVEGF receptor tyrosine kinase or Src non-receptor tyrosine kinase.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows an X-Ray Powder Diffraction Pattern of Form A of Example3.06b.

FIG. 2 shows a DSC Thermogram of Form A of Example 3.06b.

FIG. 3 shows an X-Ray Powder Diffraction Pattern of Form B of Example3.06b.

FIG. 4 shows a DSC Thermogram of Form B of Example 3.06b.

FIG. 5 shows an X-Ray Powder Diffraction Pattern of Form A of Example3.13b.

FIG. 6 shows a DSC Thermogram of Form A of Example 3.13b.

FIG. 7 shows an X-Ray Powder Diffraction Pattern of Form B of Example3.13b.

FIG. 8 shows a DSC Thermogram of Form B of Example 3.13b.

FIG. 9 shows a TGA Thermogram of Form B of Example 3.13b.

According to one aspect of the invention there is provided a chromenonederivative of the Formula I

in which:

-   R¹ is H or (1-4C)alkyl optionally substituted by 1, 2 or 3    substituents independently selected from halogeno, hydroxy or    (1-3C)alkoxy;-   R² is (1-4C)alkyl or (1-4C)alkoxy, either of which can be optionally    substituted by 1, 2 or 3 substituents independently selected from    halogeno, hydroxy, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy,    cyano, (1-3C)alkylamino or di-[(1-3C)alkyl]amino; or-   R¹ and R² together form a 3 to 8 membered nitrogen containing    heterocyclyl ring system, which optionally contains 1 or 2 further    heteroatoms selected from oxygen, nitrogen and sulphur, wherein a    ring sulphur atom is optionally oxidised to form the S-oxide(s),    said ring being optionally substituted by 1, 2 or 3 substituents    independently selected from halogeno, hydroxy, (1-3C)alkyl,    (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy, oxo,    hydroxy-(1-3C)alkyl, halogeno-(1-3C)alkyl and    (1-3C)alkoxy-(1-3C)alkyl;-   R³ is H or (1-3C)alkyl;-   R⁴ and R⁵ are independently selected from H, halogeno, (1-3C)alkyl,    (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano; or-   R⁴ and R⁵ together form a phenyl ring or a 5 or 6 membered    heterocyclyl ring or a 5 or 6 membered heteroaryl ring, wherein the    heterocyclyl or heteroaryl ring contains 1, 2 or 3 heteroatoms    selected from oxygen and nitrogen, said phenyl, heterocyclyl or    heteroaryl ring being optionally substituted by 1, 2 or 3    substituents independently selected from halogeno, (1-3C)alkyl,    (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   R⁶, R⁷ and R⁸ are independently selected from H, halogeno,    (1-3C)alkyl, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   n is 0, 1, 2, 3 or 4;-   each R⁹ group is (1-3C)alkyl; or a pharmaceutically-acceptable salt    thereof.

In this specification the generic term “(1-8C)alkyl” includes bothstraight-chain and branched-chain alkyl groups such as propyl, isopropyland tert-butyl, and also (3-8C)cycloalkyl groups such as cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, and also(3-6C)cycloalkyl-(1-2C)alkyl groups such as cyclopropylmethyl,2-cyclopropylethyl, cyclobutylmethyl, 2-cyclobutylethyl,cyclopentylmethyl, 2-cyclopentylethyl, cyclohexylmethyl and2-cyclohexylethyl. However references to individual alkyl groups such as“propyl” are specific for the straight-chain version only, references toindividual branched-chain alkyl groups such as “isopropyl” are specificfor the branched-chain version only and references to individualcycloalkyl groups such as “cyclopentyl” are specific for that 5-memberedring only. An analogous convention applies to other generic terms, forexample (1-6C)alkoxy includes (3-6C)cycloalkyloxy groups andcycloalkyl-alkoxy groups having 4 to 6 carbon atoms, for examplemethoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclobutyloxy,cyclopentyloxy, cyclohexyloxy, cyclopropylmethoxy, 2-cyclopropylethoxy,cyclobutylmethoxy, 2-cyclobutylethoxy and cyclopentylmethoxy;(1-6C)alkylamino includes (3-6C)cycloalkylamino groups andN-(cycloalkylalkyl)amino groups having 4 to 6 carbon atoms, for examplemethylamino, ethylamino, propylamino, cyclopropylamino, cyclobutylamino,cyclohexylamino, cyclopropylmethylamino, 2-cyclopropylethylamino,cyclobutylmethylamino, 2-cyclobutylethylamino andcyclopentylmethylamino; and di-[(1-6Calkyl]amino includesdi-[(3-6C)cycloalkyl]amino groups and di-[cycloalkylalkyl]amino groupsin which the cycloalkylalkyl moiety has 4 to 6 carbon atoms, for exampledimethylamino, diethylamino, dipropylamino, N-cyclopropyl-N-methylamino,N -cyclobutyl-N-methylamino, N-cyclohexyl-N-ethylamino,N-cyclopropylmethyl-N-methylamino, N-(2-cyclopropylethyl)-N-methylaminoand N-cyclopentylmethyl-N-methylamino.

A person skilled in the art will appreciate that the terms“(1-6C)alkyl”, “(1-4C)alkyl”, “(1-3C)alkyl” and “(1-2C)alkyl” that areused herein refer to any of the alkyl groups defined above thatpossesses 1 to 6, 1 to 4, 1 to 3 and 1 to 2 carbon atoms respectively.The same convention applies to other terms used herein, such as, forexample, “(1-6C)alkoxy”, “(1-4C)alkoxy”, “(1-3C)alkoxy” and“(1-2C)alkoxy”.

For the avoidance of doubt, when, as defined hereinbefore, an R⁴ and R⁵group together form a phenyl ring or a 5 or 6 membered heterocyclyl ringor a 5 or 6 membered heteroaryl ring, said ring includes the carbonatoms of the core phenyl group to which the the R⁴ and R⁵ groups areattached. For example, when the R⁴ and R⁵ group together form a phenylring, then the ring system directly attached to the N(R³) group would bea napthyl ring:

Similarly, when, as defined hereinbefore, the R⁴ and R⁵ group togetherform a pyridinyl ring, then the ring system directly attached to theN(R³) group would be a quinolinyl or isoquinolinyl ring.

It is to be understood that, insofar as certain of the compounds ofFormula I defined above may exist in optically active or racemic formsby virtue of one or more asymmetric carbon atoms, the invention includesin its definition any such optically active or racemic form whichpossesses phosphoinositide (PI) 3-kinase inhibitory activity. Thesynthesis of optically active forms may be carried out by standardtechniques of organic chemistry well known in the art, for example bysynthesis from optically active starting materials or by resolution of aracemic form. Similarly, the above-mentioned activity may be evaluatedusing the standard laboratory techniques.

A particular enantiomer of the compounds described herein may be moreactive that other enantiomers of the compound. For example, the (+)enantiomer of the title compound of Example 3.06 (i.e. the compound ofExample 3.06a, where (+) signifies the optical rotation measured usingthe conditions described in Example 3.06a) is the enantiomer having theweaker activity. For the avoidance of doubt, the chiral centre inquestion is the carbon atom to which the groups methyl and—N(R³)phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) are attached.

Accordingly, in a further aspect of the invention, there is provided achromenone derivative of the Formula I, or a pharmaceutically-acceptablesalt thereof, wherein the chiral centre to which the groups methyl and—N(R³)phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) are attached is in the(R)-stereochemical configuration. In a further aspect of the invention,there is provided a chromenone derivative of the Formula I, or apharmaceutically-acceptable salt thereof, wherein the chiral centre towhich the groups methyl and —N(R³)phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) areattached is in the (S)-stereochemical configuration.

According to a further aspect of the invention there is provided achromenone derivative of the Formula I, or a pharmaceutically-acceptablesalt thereof, which is a single enantiomer being in an enantiomericexcess (% ee) of ≧95, ≧98% or ≧99%. In one embodiment of this aspect ofthe invention, the chiral centre to which the groups methyl and—N(R³)phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) are attached is in the(R)-stereochemical configuration. In a further embodiment of this aspectof the invention, the chiral centre to which the groups methyl and—N(R³)phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) are attached is in the(S)-stereochemical configuration.

According to a further aspect of the invention there is provided apharmaceutical composition, which comprises a chromenone derivative ofthe Formula I, which is a single enantiomer being in an enantiomericexcess (% ee) of ≧95, ≧98% or ≧99% or a pharmaceutically-acceptable saltthereof, in association with a pharmaceutically-acceptable diluent orcarrier. Conveniently, the single enantiomer is present in anenantiomeric excess (% ee) of ≧99%. In one embodiment of this aspect ofthe invention, the chiral centre to which the groups methyl and—N(R³)phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) are attached is in the(R)-stereochemical configuration. In a further embodiment of this aspectof the invention, the chiral centre to which the groups methyl and—N(R³)phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) are attached is in the(S)-stereochemical configuration.

Some compounds of formula (I) may exhibit polymorphism. It is to beunderstood that the present invention encompasses any polymorphic form,or mixtures thereof, which form possesses properties useful in theinhibition of phosphoinositide (PI) 3-kinase activity, it being wellknown in the art how to determine efficacy of a polymorphic form for theinhibition of phosphoinositide (PI) 3-kinase activity by the standardtests described hereinafter.

It is generally known that crystalline materials may be analysed usingconventional techniques such as X-Ray Powder Diffraction (hereinafterXRPD) analysis, Differential Scanning Calorimetry (hereinafter DSC),Thermal Gravimetric Analysis (hereinafter TGA), Diffuse ReflectanceInfrared Fourier Transform (DRIFT) spectroscopy, Near Infrared (NIR)spectroscopy, solution and/or solid state nuclear magnetic resonancespectroscopy. The water content of such crystalline materials may bedetermined by Karl Fischer analysis.

As an example, the compound of Example 3.06b exhibits polymorphism andtwo crystalline forms have been identified Accordingly, a further aspectof the invention is Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,where the (−)—in the chemical name signifies the optical rotationmeasured using the conditions described in Example 3.06b.

Accordingly, a further aspect of the invention is Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide, which has anX-ray powder diffraction pattern with at least one specific peak atabout 2-theta=7.9°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at about 2-theta=16.7°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at about 2-theta=7.9° and 16.7°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks atabout 2-theta=7.9, 16.7, 20.3, 19.3, 13.2, 7.2, 19.5, 17.9, 23.0, 5.0°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern substantially the same asthe X-ray powder diffraction pattern shown in FIG. 1.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=7.9° plus or minus 0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=16.7° plus or minus 0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=7.9° and 16.7° wherein said values may be plus or minus0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=7.9, 16.7, 20.3, 19.3, 13.2, 7.2, 19.5, 17.9, 23.0, 5.0°.wherein said values may be plus or minus 0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=7.90°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=16.7°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=7.9° and 16.7°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=7.9, 16.7, 20.3, 19.3, 13.2, 7.2, 19.5, 17.9, 23.0, 5.0°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form A of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern as shown in FIG. 1.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at about 2-theta=20.7°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at about 2-theta=13.8°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at about 2-theta=20.7° and 13.8°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks atabout 2-theta=20.7, 13.8, 21.5, 19.6, 12.8, 15.4, 10.7, 8.5, 22.4°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern substantially the same asthe X-ray powder diffraction pattern shown in FIG. 3.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=20.7° plus or minus 0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=13.8° plus or minus 0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=20.7° and 13.8° wherein said values may be plus orminus 0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=20.7, 13.8, 21.5, 19.6, 12.8, 15.4, 10.7, 8.5, 22.4° whereinsaid values may be plus or minus 0.5° 2-theta.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=20.7°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=13.8°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=20.7° and 13.8°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=20.7, 13.8, 21.5, 19.6, 12.8, 15.4, 10.7, 8.5, 22.4°.

According to a further aspect of the present invention, there isprovided a crystalline form, Form B of(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern as shown in FIG. 3.

A further example of a compound exhibiting polymorphism is the compoundof Example 3.13b.

Accordingly, a further aspect of the invention is Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,where the (−)- in the chemical name signifies the optical rotationmeasured using the conditions described in Example 3.13b.

Accordingly, a further aspect of the invention is Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at about 2-theta=20.0°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at about 2-theta=18.0°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at about 2-theta=20.0° and 18.0°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks atabout 2-theta=20.0, 18.0, 14.0, 19.4, 23.2, 23.8, 10.8, 19.1, 11.2,27.8°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern substantially the same asthe X-ray powder diffraction pattern shown in FIG. 5.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=20.0° plus or minus 0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=18.0° plus or minus 0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=20.0° and 18.0° wherein said values may be plus orminus 0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=20.0, 18.0, 14.0, 19.4, 23.2, 23.8, 10.8, 19.1, 11.2, 27.8°wherein said values may be plus or minus 0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=20.0°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=18.0°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=20.0° and 18.0°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=20.0, 18.0, 14.0, 19.4, 23.2, 23.8, 10.8, 19.1, 11.2, 27.8°.

According to a further aspect of the invention there is provided acrystalline form, Form A of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern as shown in FIG. 5.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at about 2-theta=6.2°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at about 2-theta=7.0°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at about 2-theta=6.2° and 7.0°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks atabout 2-theta=6.2, 7.0, 10.3, 22.4, 15.9, 20.4, 27.2, 12.4, 18.7, 12.8°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern substantially the same asthe X-ray powder diffraction pattern shown in FIG. 7.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=6.2° plus or minus 0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=7.0° plus or minus 0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=6.2° and 7.0° wherein said values may be plus or minus0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=6.2, 7.0, 10.3, 22.4, 15.9, 20.4, 27.2, 12.4, 18.7, 12.8°wherein said values may be plus or minus 0.5° 2-theta.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=6.2°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least one specificpeak at 2-theta=7.0°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with at least two specificpeaks at 2-theta=6.2° and 7.0°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern with specific peaks at2-theta=6.2, 7.0, 10.3, 22.4, 15.9, 20.4, 27.2, 12.4, 18.7, 12.8°.

According to a further aspect of the invention there is provided acrystalline form, Form B of(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide,which has an X-ray powder diffraction pattern as shown in FIG. 7.

It will be understood that 2-theta values of the X-ray powderdiffraction patterns may vary slightly from one machine to another orfrom one sample to another, and so the values quoted are not to beconstrued as absolute.

It is known that an X-ray powder diffraction pattern may be obtainedwhich has one or more measurement errors depending on measurementconditions (such as equipment or machine used). In particular, it isgenerally known that intensities in an X-ray powder diffraction patternmay fluctuate depending on measurement conditions. Therefore it shouldbe understood that the crystalline Forms of the present inventiondescribed above, unless otherwise stated, are not limited to thecrystals that provide X-ray powder diffraction patterns identical to theX-ray powder diffraction pattern shown in FIGS. 1, 3, 5 and 7 and anycrystals providing X-ray powder diffraction patterns substantially thesame as those shown in these Figures fall within the scope of thepresent invention. A person skilled in the art of X-ray powderdiffraction is able to judge the substantial identity of X-ray powderdiffraction patterns.

Persons skilled in the art of X-ray powder diffraction will also realisethat the relative intensity of peaks can be affected by, for example,grains above 30 microns in size and non-unitary aspect ratios, which mayaffect analysis of samples. The skilled person will also realise thatthe position of reflections can be affected by the precise height atwhich the sample sits in the diffractometer and the zero calibration ofthe diffractometer. The surface planarity of the sample may also have asmall effect. Hence the diffraction pattern data presented are not to betaken as absolute values (see Jenkins, R & Snyder, R. L. ‘Introductionto X-Ray Powder Diffractometry’ John Wiley & Sons 1996; Bunn, C. W.(1948), Chemical Crystallography, Clarendon Press, London; Klug, H. P. &Alexander, L. E. (1974), X-Ray Diffraction Procedures).

Generally, a measurement error of a diffraction angle in an X-ray powderdiffractogram is approximately plus or minus 0.5° 2-theta, and suchdegree of a measurement error should be taken into account whenconsidering the X-ray powder diffraction data. Furthermore, it should beunderstood that intensities might fluctuate depending on experimentalconditions and sample preparation (preferred orientation).

Particular compounds of the invention are each of the Examples andpharmaceutically-acceptable salt(s) thereof, each of which provides afurther independent aspect of the invention.

According to a further aspect of the invention there is provided achromenone derivative of the Formula I, which is obtainable by followingany of the Examples as disclosed herein.

A further feature is any of the scopes defined herein with the provisothat specific Examples, such as Example 1.00, 2.00, 3.00, 4.00 etc. areindividually disclaimed.

It is to be understood that certain compounds of Formula I defined abovemay exhibit the phenomenon of tautomerism. It is to be understood thatthe present invention includes in its definition any such tautomericform, or a mixture thereof, which possesses phosphoinositide (PI)3-kinase inhibitory activity and is not to be limited merely to any onetautomeric form utilised within the formulae drawings or named in theExamples. In general, just one of any such tautomeric forms is named inthe Examples that follow hereinafter or is presented in any relevantformulae drawings that follow hereinafter.

Suitable values for the generic radicals referred to above include thoseset out below.

A suitable value for the 3 to 8 membered nitrogen containingheterocyclyl ring system formed by the R¹ and R² groups of Formula I is,for example, a nitrogen containing non-aromatic saturated or partiallysaturated 3 to 8 membered ring, which optionally contains 1 or 2 furtherheteroatoms selected from oxygen, nitrogen and sulphur, wherein a ringsulphur atom is optionally oxidised to form the S-oxide(s). Suitableexamples include azepanyl, oxazepanyl, aziridinyl, azetidinyl,pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, pyrazolinyl,pyrazolidinyl, morpholinyl, thiomorpholinyl, tetrahydro-1,4-thiazinyl,1,1-dioxotetrahydro-1,4-thiazinyl, piperidinyl, homopiperidinyl,piperazinyl, homopiperazinyl, dihydropyridinyl, tetrahydropyridinyl,dihydropyrimidinyl or tetrahydropyrimidinyl. In a particular group ofcompounds, particular examples of the heterocyclyl ring includeazepanyl, oxazepanyl, azetidinyl, pyrrolidinyl, morpholinyl,thiomorpholinyl, piperidinyl, piperazinyl, and especially azepan-1-yl,1,4-oxazepan-4-yl, azetidin-1-yl, pyrrolidine-1-yl, morpholin-4-yl,thiomorpholin-4-yl, piperidin-1-yl and piperazine-1-yl.

A suitable value for the 5 to 6 membered heterocyclyl ring formed by theR⁴ and R⁵ groups of Formula I is, for example, a non-aromatic saturatedor partially saturated 5 or 6 membered ring, which contains 1, 2 or 3heteroatoms selected from oxygen and nitrogen. Suitable examples includetetrahydrofuranyl, tetrahydropyranyl, pyrrolinyl, pyrrolidinyl,imidazolinyl, imidazolidinyl, pyrazolinyl, pyrazolidinyl, morpholinyl,piperidinyl, homopiperidinyl, piperazinyl, homopiperazinyl,dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl ortetrahydropyrimidinyl.

A suitable value for the 5 to 6 membered heteroaryl ring formed by theR⁴ and R⁵ groups of Formula I is, for example, an aromatic 5- or6-membered monocyclic ring with 1, 2 or 3 ring heteroatoms selected fromoxygen and nitrogen. Suitable examples include furanyl, pyrrolyl,oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, oxadiazolyl, triazolyl,pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl or 1,3,5-triazinyl.

Suitable values for any of the ‘R’ groups (R¹ to R⁹), include, forexample:—

-   for halogeno fluoro, chloro, bromo and iodo;-   for (1-8C)alkyl: methyl, ethyl, propyl, isopropyl, tert-butyl,    cyclobutyl, cyclohexyl, cyclohexylmethyl and 2-cyclopropylethyl;-   for (2-8C)alkenyl: vinyl, isopropenyl, allyl and but-2-enyl;-   for (2-8C)alkynyl: ethynyl, 2-propynyl and but-2-ynyl;-   for (1-6C)alkoxy: methoxy, ethoxy, propoxy, isopropoxy and butoxy;-   for (1-6C)alkylamino: methylamino, ethylamino, propylamino,    isopropylamino and butylamino;-   for di-[(1-6C)alkyl]amino: dimethylamino, diethylamino,    N-ethyl-N-methylamino and diisopropylamino;-   for halogeno-(1-6C)alkyl: chloromethyl, 2-fluoroethyl,    2-chloroethyl, 1-chloroethyl, 2,2-difluoroethyl,    2,2,2-trifluoroethyl, 3-fluoropropyl, 3-chloropropyl,    3,3-difluoropropyl and 3,3,3-trifluoropropyl;-   for hydroxy-(1-6C)alkyl: hydroxymethyl, 2-hydroxyethyl,    1-hydroxyethyl and 3-hydroxypropyl; and-   for (1-6C)alkoxy-(1-6C)alkyl: methoxymethyl, ethoxymethyl,    1-methoxyethyl, 2-methoxyethyl, 2-ethoxyethyl and 3-methoxypropyl.

A suitable pharmaceutically-acceptable salt of a compound of the FormulaI is, for example, an acid-addition salt of a compound of the Formula I,for example an acid-addition salt with an inorganic or organic acid suchas hydrochloric, hydrobromic, sulphuric, trifluoroacetic or citric acid;or, for example, a salt of a compound of the Formula I which issufficiently acidic, for example an alkali or alkaline earth metal saltsuch as a calcium or magnesium salt, or an ammonium salt, or a salt withan organic base such as methylamine, dimethylamine, trimethylamine,piperidine, morpholine or tris-(2-hydroxyethyl)amine. A further suitablepharmaceutically-acceptable salt of a compound of the Formula I is, forexample, a salt formed within the human or animal body afteradministration of a compound of the Formula I.

It is further to be understood that a suitablepharmaceutically-acceptable solvate of a compound of the Formula I alsoforms an aspect of the present invention. A suitablepharmaceutically-acceptable solvate is, for example, a hydrate such as ahemi-hydrate, a mono-hydrate, a di-hydrate or a tri-hydrate or analternative quantity thereof.

It is further to be understood that a suitablepharmaceutically-acceptable pro-drug of a compound of the Formula I alsoforms an aspect of the present invention. Accordingly, the compounds ofthe invention may be administered in the form of a pro-drug, that is acompound that is broken down in the human or animal body to release acompound of the invention. A pro-drug may be used to alter the physicalproperties and/or the pharmacokinetic properties of a compound of theinvention. A pro-drug can be formed when the compound of the inventioncontains a suitable group or substituent to which a property-modifyinggroup can be attached. Examples of pro-drugs include in vivo cleavableester derivatives that may be formed at a carboxy group or a hydroxygroup in a compound of the Formula I and in vivo cleavable amidederivatives that may be formed at a carboxy group or an amino group in acompound of the Formula I.

Accordingly, the present invention includes those compounds of theFormula I as defined hereinbefore when made available by organicsynthesis and when made available within the human or animal body by wayof cleavage of a pro-drug thereof. Accordingly, the present inventionincludes those compounds of the Formula I that are produced by organicsynthetic means and also such compounds that are produced in the humanor animal body by way of metabolism of a precursor compound, that is acompound of the Formula I may be a synthetically-produced compound or ametabolically-produced compound.

A suitable pharmaceutically-acceptable pro-drug of a compound of theFormula I is one that is based on reasonable medical judgement as beingsuitable for administration to the human or animal body withoutundesirable pharmacological activities and without undue toxicity.

Various forms of pro-drug have been described, for example in thefollowing documents:—

-   a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder,    et al. (Academic Press, 1985);-   b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier, 1985);-   c) A Textbook of Drug Design and Development, edited by    Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and    Application of Pro-drugs”, by H. Bundgaard p. 113-191 (1991);-   d) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);-   e) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285    (1988);-   f) N. Kakeya, et al., Chem. Pharm. Bull., 32, 692 (1984);-   g) T. Higuchi and V. Stella, “Pro-Drugs as Novel Delivery Systems”,    A.C.S. Symposium Series, Volume 14; and-   h) E. Roche (editor), “Bioreversible Carriers in Drug Design”,    Pergamon Press, 1987.

A suitable pharmaceutically-acceptable pro-drug of a compound of theFormula I that possesses a carboxy group is, for example, an in vivocleavable ester thereof. An in vivo cleavable ester of a compound of theFormula I containing a carboxy group is, for example, apharmaceutically-acceptable ester which is cleaved in the human oranimal body to produce the parent acid. Suitablepharmaceutically-acceptable esters for carboxy include (1-6C)alkylesters such as methyl, ethyl and tert-butyl, (1-6C)alkoxymethyl esterssuch as methoxymethyl esters, (1-6C)alkanoyloxymethyl esters such aspivaloyloxymethyl esters, 3-phthalidyl esters,(3-8C)cycloalkylcarbonyloxy-(1-6C)alkyl esters such ascyclopentylcarbonyloxymethyl and 1-cyclohexylcarbonyloxyethyl esters,2-oxo-1,3-dioxolenylmethyl esters such as5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl esters and(1-6C)alkoxycarbonyloxy-(1-6C)alkyl esters such asmethoxycarbonyloxymethyl and 1-methoxycarbonyloxyethyl esters.

A suitable pharmaceutically-acceptable pro-drug of a compound of theFormula I that possesses a hydroxy group is, for example, an in vivocleavable ester or ether thereof. An in vivo cleavable ester or ether ofa compound of the Formula I containing a hydroxy group is, for example,a pharmaceutically-acceptable ester or ether which is cleaved in thehuman or animal body to produce the parent hydroxy compound. Suitablepharmaceutically-acceptable ester forming groups for a hydroxy groupinclude inorganic esters such as phosphate esters (includingphosphoramidic cyclic esters). Further suitablepharmaceutically-acceptable ester forming groups for a hydroxy groupinclude (1-10C)alkanoyl groups such as acetyl, benzoyl, phenylacetyl andsubstituted benzoyl and phenylacetyl groups, (1-10C)alkoxycarbonylgroups such as ethoxycarbonyl, N,N-[di-(1-4C)alkyl]carbamoyl,2-dialkylaminoacetyl and 2-carboxyacetyl groups. Examples of ringsubstituents on the phenylacetyl and benzoyl groups include aminomethyl,N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl,piperazin-1-ylmethyl and 4-(1-4C)alkylpiperazin-1-ylmethyl. Suitablepharmaceutically-acceptable ether forming groups for a hydroxy groupinclude α-acyloxyalkyl groups such as acetoxymethyl andpivaloyloxymethyl groups.

A suitable pharmaceutically-acceptable pro-drug of a compound of theFormula I that possesses a carboxy group is, for example, an in vivocleavable amide thereof, for example an amide formed with an amine suchas ammonia, a (1-4C)alkylamine such as methylamine, adi-(1-4C)alkylamine such as dimethylamine, N-ethyl-N-methylamine ordiethylamine, a (1-4C)alkoxy-(2-4C)alkylamine such as2-methoxyethylamine, a phenyl-(1-4C)alkylamine such as benzylamine andamino acids such as glycine or an ester thereof.

A suitable pharmaceutically-acceptable pro-drug of a compound of theFormula I that possesses an amino group is, for example, an in vivocleavable amide derivative thereof. Suitable pharmaceutically-acceptableamides from an amino group include, for example an amide formed with(1-10C)alkanoyl groups such as an acetyl, benzoyl, phenylacetyl andsubstituted benzoyl and phenylacetyl groups. Examples of ringsubstituents on the phenylacetyl and benzoyl groups include aminomethyl,N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl,piperazin-1-ylmethyl and 4-(1-4C)alkylpiperazin-1-ylmethyl.

The in vivo effects of a compound of the Formula I may be exerted inpart by one or more metabolites that are formed within the human oranimal body after administration of a compound of the Formula I. Asstated hereinbefore, the in vivo effects of a compound of the Formula Imay also be exerted by way of metabolism of a precursor compound (apro-drug).

For the avoidance of doubt it is to be understood that where in thisspecification a group is qualified by ‘hereinbefore defined’ or ‘definedhereinbefore’ the said group encompasses the first occurring andbroadest definition as well as each and all of the particulardefinitions for that group.

Particular novel compounds of the invention include, for example,chromenone derivatives of the Formula I, or pharmaceutically-acceptablesalts thereof, wherein, unless otherwise stated, each of R¹, R², R³, R⁴,R⁵, R⁶, R⁷, R⁸, n and R⁹ has any of the meanings defined hereinbefore orin paragraphs (a) to (ii) hereinafter:—

-   (a) R¹ is H or (1-4C)alkyl;-   (b) R¹ is H;-   (c) R¹ is (1-4C)alkyl;-   (d) R¹ is methyl or ethyl;-   (e) R¹ is H, methyl or ethyl;-   (f) R¹ is methyl;-   (g) R² is (1-4C)alkyl optionally substituted by halogeno, hydroxy,    (1-3C)alkoxy, cyano, (1-3C)alkylamino or di-[(1-3C)alkyl]amino;-   (h) R² is (1-3C)alkyl optionally substituted by halogeno, hydroxy,    (1-3C)alkoxy or di-[(1-3C)alkyl]amino;-   (i) R² is (1-3C)alkyl optionally substituted by halogeno, hydroxy,    methoxy or N,N-dimethylamino;-   (j) R² is methyl, ethyl, propyl, 2-fluoroethyl, 2-hydroxyethyl,    2-methoxyethyl, 3-methoxypropyl, cyclopropylmethyl or    1-(N,N-dimethylamino)ethyl;-   (k) R² is methyl;-   (l) R¹ and R² are both methyl;-   (m) R¹ and R² together form a 4 to 7 membered nitrogen containing    heterocyclyl ring system, which optionally contains 1 further    heteroatom selected from oxygen, nitrogen and sulphur, wherein a    ring sulphur atom is optionally oxidised to form the S-oxide(s),    said ring being optionally substituted by halogeno, hydroxy,    (1-3C)alkyl, (1-3C)alkoxy or hydroxy-(1-3C)alkyl;-   (n) R¹ and R² together form a nitrogen containing heterocyclyl ring    system, selected from azepanyl, oxazepanyl, azetidinyl,    pyrrolidinyl, morpholinyl, thiomorpholinyl, piperidinyl and    piperazinyl, said ring being optionally substituted by halogeno,    hydroxy, (1-3C)alkyl, (1-3C)alkoxy or hydroxy-(1-3C)alkyl;-   (o) R¹ and R² together form a nitrogen containing heterocyclyl ring    system, selected from azepan-1-yl, 1,4-oxazepan-4-yl, azetidin-1-yl,    pyrrolidine-1-yl, morpholin-4-yl, thiomorpholin-4-yl,    piperidine-1-yl and piperazine-1-yl, said ring being optionally    substituted by halogeno, hydroxy, (1-3C)alkyl, (1-3C)alkoxy or    hydroxy-(1-3C)alkyl;-   (p) R³ is H or methyl;-   (q) R³ is H;-   (r) R³ is methyl;-   (s) R⁴ and R⁵ are independently selected from H, halogeno,    (1-3C)alkyl, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   (t) R⁴ and R⁵ are independently selected from H, fluoro, chloro,    methyl, ethynyl, methoxy and cyano;-   (u) R⁴ and R⁵ are independently selected from H or halogeno;-   (v) R⁴ and R⁵ are independently selected from H, fluoro or chloro;-   (w) R⁶, R⁷ and R⁸ are independently selected from H, fluoro, chloro,    methyl, ethynyl, methoxy and cyano;-   (x) R⁶, R⁷ and R⁸ are independently selected from H or halogeno;-   (y) R⁶ is H and R⁷ and R⁸ are independently selected from H, fluoro    or chloro;-   (z) R⁶ and R⁸ are H and R⁴, R⁵ and R⁷ are halogeno;-   (aa) R⁶ and R⁸ are H and R⁴, R⁵ and R⁷ are fluoro;-   (bb) R⁴, R⁶ and R⁸ are H and R⁵ and R⁷ are halogeno;-   (cc) R⁴, R⁶ and R⁸ are H and R⁵ and R⁷ are fluoro;-   (dd) n is 0;-   (ee) n is 0 or 1;-   (ff) n is 1;-   (gg) R⁹ is methyl or ethyl;-   (hh) R⁹ is methyl; or-   (ii) n is 1 and R⁹ is a methyl group located in the 2-position of    the morpholine ring.

A particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is H or (1-4C)alkyl;-   R² is (1-4C)alkyl optionally substituted by halogeno, hydroxy,    (1-3C)alkoxy, cyano, (1-3C)alkylamino or di-[(1-3C)alkyl]amino; or-   R¹ and R² together form a 4 to 7 membered nitrogen containing    heterocyclyl ring system, which optionally contains 1 further    heteroatom selected from oxygen, nitrogen and sulphur, wherein a    ring sulphur atom is optionally oxidised to form the S-oxide(s),    said ring being optionally substituted by halogeno, hydroxy,    (1-3C)alkyl, (1-3C)alkoxy or hydroxy-(1-3C)alkyl;-   R³ is H or methyl;-   R⁴ and R⁵ are independently selected from H, halogeno, (1-3C)alkyl,    (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   R⁶, R⁷ and R⁸ are independently selected from H, halogeno,    (1-3C)alkyl, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   n is 0; or a pharmaceutically-acceptable salt thereof.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is H, methyl or ethyl;-   R² is (1-4C)alkyl optionally substituted by halogeno, hydroxy,    (1-3C)alkoxy, cyano, (1-3C)alkylamino or di-[(1-3C)alkyl]amino; or-   R¹ and R² together form a nitrogen containing heterocyclyl ring    system, selected from azepanyl, oxazepanyl, azetidinyl,    pyrrolidinyl, morpholinyl, thiomorpholinyl, piperidinyl and    piperazinyl, said ring being optionally substituted by halogeno,    hydroxy, (1-3C)alkyl, (1-3C)alkoxy or hydroxy-(1-3C)alkyl;-   ³ is H;-   R⁴ and R⁵ are independently selected from H, fluoro or chloro;-   R⁶, R⁷ and R⁸ are independently selected from H or halogeno;-   n is 0; or a pharmaceutically-acceptable salt thereof.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ and R² are suitably as defined in any of paragraphs (a) to (g)    and (m) to (o) above;-   R³ is suitably as defined in any one of paragraphs (p) to (q) above;-   R⁴ and R⁵ are suitably as defined in any one of paragraphs (s), (u)    to (v) above and is particularly as defined in any one of    paragraphs (u) to (v) above;-   R⁶, R⁷ and R⁸ are suitably as defined in any one of paragraphs (x)    to (y) above; and-   n is suitably as defined in paragraph (dd) above.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above or pharmaceutically-acceptable salt(s)thereof, wherein:—

-   R¹ and R² are suitably as defined in any of paragraphs (a) to (o)    above, particularly as defined in paragraph (1) above;-   R³ is suitably as defined in any one of paragraphs (p) to (r) above;-   R⁴ and R⁵ are suitably as defined in any one of paragraphs (s)    to (v) above and is particularly as defined in paragraph (v) above;-   R⁶, R⁷ and R⁸ are suitably as defined in any one of paragraphs (w)    to (cc) above and is particularly as defined in any one of    paragraphs (z) to (cc) above; and-   n and R⁹ are suitably as defined in any one of paragraphs (dd)    to (ii) above.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is H, methyl or ethyl;-   R² is methyl, ethyl, cyclopropylmethyl, 2-fluoroethyl,    2-hydroxyethyl, 2-methoxyethyl, 3-methoxypropyl or    2-(dimethylamino)ethyl; or-   R¹ and R² together form an optionally substituted nitrogen    containing heterocyclyl ring system, selected from azepan-1-yl,    1,4-oxazepan-4-yl, azetidin-1-yl, 3-fluoroazetidin-1-yl,    3-hydroxyazetidin-1-yl, pyrrolidin-1-yl,    (2R)-2-(hydroxymethyl)pyrrolidin-1-yl,    (2S)-2-(hydroxymethyl)pyrrolidin-1-yl, morpholin-4-yl,    thiomorpholin-4-yl, piperidin-1-yl, 4-hydroxypiperidin-1-yl,    4-methoxypiperidin-1-yl, piperazin-1-yl or 4-methylpiperazin-1-yl;-   R³ is H or methyl;-   R⁴ and R⁵ are independently selected from H, fluoro or chloro;-   R⁶, R⁷ and R⁸ are independently selected from H, fluoro or chloro;-   n is 0; or a pharmaceutically-acceptable salt thereof.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is methyl;-   R² is methyl; or-   R¹ and R² together form a pyrrolidin-1-yl ring;-   R³ is H;-   R⁴ and R⁵ are independently selected from H or fluoro;-   R⁶ is H;-   R⁷ and R⁸ are independently selected from H, fluoro or chloro;-   n is 0; or a pharmaceutically-acceptable salt thereof

A particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is H or (1-4C)alkyl;-   R² is (1-4C)alkyl optionally substituted by halogeno, hydroxy,    (1-3C)alkoxy, cyano, (1-3C)alkylamino or di-[(1-3C)alkyl]amino; or-   R¹ and R² together form a 4 to 7 membered nitrogen containing    heterocyclyl ring system, which optionally contains 1 further    heteroatom selected from oxygen, nitrogen and sulphur, wherein a    ring sulphur atom is optionally oxidised to form the S-oxide(s),    said ring being optionally substituted by halogeno, hydroxy,    (1-3C)alkyl, (1-3C)alkoxy or hydroxy-(1-3C)alkyl;-   R³ is H or methyl;-   R⁴ and R⁵ are independently selected from H, halogeno, (1-3C)alkyl,    (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   R⁶, R⁷ and R⁸ are independently selected from H, halogeno,    (1-3C)alkyl, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   n is 0 or 1;-   each R⁹ group is methyl; or a pharmaceutically-acceptable salt    thereof.

A particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is methyl;-   R² is methyl or (2-hydroxy)ethyl; or-   R¹ and R² together form a 6 membered nitrogen containing    heterocyclyl ring system, said ring system being optionally    substituted by hydroxy;-   R³ is H or methyl;-   R⁴ and R⁵ are independently selected from H or halogeno;-   R⁶, R⁷ and R⁸ are independently selected from H or halogeno;-   n is 1;-   R⁹ is a methyl group located in the 2-position of the morpholine    ring; or a pharmaceutically-acceptable salt thereof.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is H, methyl or ethyl;-   R² is (1-3C)alkyl optionally substituted by halogeno, hydroxy,    (1-3C)alkoxy, cyano, (1-3C)alkylamino or di-[(1-3C)alkyl]amino; or-   R¹ and R² together form a nitrogen containing heterocyclyl ring    system, selected from azepanyl, oxazepanyl, azetidinyl,    pyrrolidinyl, morpholinyl, thiomorpholinyl, piperidinyl and    piperazinyl, said ring being optionally substituted by halogeno,    hydroxy, (1-3C)alkyl, (1-3C)alkoxy or hydroxy-(1-3C)alkyl;-   R³ is H;-   R⁴ and R⁵ are independently selected from H, halogeno, (1-3C)alkyl,    (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   R⁶, R⁷ and R⁸ are independently selected from H, halogeno,    (1-3C)alkyl, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy and cyano;-   n is 0 or 1;-   each R⁹ group is (1-3C)alkyl; or a pharmaceutically-acceptable salt    thereof.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is H, methyl or ethyl;-   R² is methyl, ethyl, cyclopropylmethyl, 2-fluoroethyl,    2-hydroxyethyl, 2-methoxyethyl, 3-methoxypropyl or    2-(dimethylamino)ethyl; or-   R¹ and R² together form an optionally substituted nitrogen    containing heterocyclyl ring system, selected from azepan-1-yl,    1,4-oxazepan-4-yl, azetidin-1-yl, 3-fluoroazetidin-1-yl,    3-hydroxyazetidin-1-yl, pyrrolidin-1-yl,    (2R)-2-(hydroxymethyl)pyrrolidin-1-yl,    (2S)-2-(hydroxymethyl)pyrrolidin-1-yl, morpholin-4-yl,    thiomorpholin-4-yl, piperidin-1-yl, 4-hydroxypiperidin-1-yl,    4-methoxypiperidin-1-yl, piperazin-1-yl or 4-methylpiperazin-1-yl;-   R³ is H or methyl;-   R⁴ and R⁵ are independently selected from H, fluoro, chloro, methyl,    ethynyl, methoxy and cyano;-   R⁶, R⁷ and R⁸ are independently selected from H, fluoro, chloro,    methyl, ethynyl, methoxy and cyano;-   n is 0 or 1;-   R⁹ is methyl; or a pharmaceutically-acceptable salt thereof.

A further particular group of compounds of the invention are chromenonederivatives of Formula I above wherein:—

-   R¹ is methyl;-   R² is methyl;-   R³ is H or methyl;-   R⁴ and R⁵ are independently selected from H or fluoro;-   R⁶ is H;-   R⁷ and R⁸ are independently selected from H or fluoro;-   n is 0; or a pharmaceutically-acceptable salt thereof

Particular compounds of the invention are, for example, the chromenonederivatives of the Formula I that are disclosed within the Examples thatare set out hereinafter.

For example, a particular compound of the invention is a chromenonederivative of the Formula I selected from any one of the following:—

-   N-(2-(dimethylamino)ethyl)-8-(1-(4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N-(2-(dimethylamino)ethyl)-8-(1-(3-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-8-(1-(phenylamino)ethyl)-4H-chromene-6-carboxamide;-   8-(1-(3-chloro-2-fluorophenylamino)ethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N-(2-(dimethylamino)ethyl)-8-(1-(4-fluorophenylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3-chloro-4-fluorophenylamino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3-chloro-4-fluorophenylamino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3-chloro-2-fluorophenylamino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3-chloro-2-fluorophenylamino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)    ethyl)-6-(morpholine-4-carbonyl)-2-morpholino-4H-chromen-4-one;-   N-(2-(dimethylamino)ethyl)-8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3,4-difluorophenyl)(methyl)amino)ethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N-(2-(dimethylamino)ethyl)-8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3-chloro-2-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3-chloro-4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3-chloro-4-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3-chlorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(2,3-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N,N-dimethyl-2-morpholino-4-oxo-8-(1-(3,4,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;-   8-(1-(3-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide-   8-(1-(3,5-difluorophenylamino)ethyl)-N,N-diethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-6-(4-methylpiperazine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-6-(piperazine-1-carbonyl)-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-6-(thiomorpholine-4-carbonyl)-4H-chromen-4-one;-   6-(azepane-1-carbonyl)-8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   6-(azetidine-1-carbonyl)-8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-6-(piperidine-1-carbonyl)-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-ethyl-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-6-(3-hydroxyazetidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-6-(3-fluoroazetidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-(2-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-(2-methoxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-N-propyl-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-ethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-(2-fluoroethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-(3-methoxypropyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-6-((R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-6-((S)-2-(hydroxymethyl)pyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N-(cyclopropylmethyl)-8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N-(2-methoxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-6-(1,4-oxazepane-4-carbonyl)-4H-chromen-4-one;    and-   8-(1-(3,5-difluorophenylamino)ethyl)-6-(4-methoxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;    or a pharmaceutically-acceptable salt thereof.

According to a further aspect of the invention, a particular compound ofthe invention is a chromenone derivative of the Formula I selected fromany one of the following:—

-   8-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide,-   8-(1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;-   N,N-dimethyl-2-morpholino-4-oxo-8-((1R)-1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;-   N,N-dimethyl-2-morpholino-4-oxo-8-((1S)-1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;-   8-(1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3,5-difluorophenyl)(methyl)    amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)    amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)    amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-((R)-2-methylmorpholino)-4H-chromen-4-one;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-((R)-2-methylmorpholino)-4H-chromen-4-one;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-((R)-2-methylmorpholino)-4H-chromen-4-one;-   8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;    and-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;    or a pharmaceutically-acceptable salt thereof.

According to a yet further aspect of the invention, a particularcompound of the invention is a chromenone derivative of the Formula Iselected from any one of the following:—

-   8-((1R)-1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N,N-dimethyl-2-morpholino-4-oxo-8-((1R)-1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;-   8-((1R)-1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)    amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1R)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-((R)-2-methylmorpholino)-4H-chromen-4-one;    and-   8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;    or a pharmaceutically-acceptable salt thereof.

According to a yet further aspect of the invention, a particularcompound of the invention is a chromenone derivative of the Formula Iselected from any one of the following:—

-   8-((1S)-1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   N,N-dimethyl-2-morpholino-4-oxo-8-((1S)-1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;-   8-((1S)-1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)    amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one;-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;-   8-((1S)-1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-((R)-2-methylmorpholino)-4H-chromen-4-one;    and-   8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide;    or a pharmaceutically-acceptable salt thereof.

According to a further aspect of the invention, a particular compound ofthe invention is the compound of Example 3.06b; or apharmaceutically-acceptable salt thereof.

According to a further aspect of the invention, a particular compound ofthe invention is(−)-8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;or a pharmaceutically-acceptable salt thereof, where the (−)- in thechemical name signifies the optical rotation measured using theconditions described in Example 3.06b.

According to a further aspect of the invention, a particular compound ofthe invention is8-((1R)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;or a pharmaceutically-acceptable salt thereof.

According to a further aspect of the invention, a particular compound ofthe invention is8-((1S)-1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide;or a pharmaceutically-acceptable salt thereof.

According to a further aspect of the invention, a particular compound ofthe invention is the compound of Example 3.13b; or apharmaceutically-acceptable salt thereof.

According to a further aspect of the invention, a particular compound ofthe invention is(−)-N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;or a pharmaceutically-acceptable salt thereof, where the (−)- in thechemical name signifies the optical rotation measured using theconditions described in Example 3.13b.

According to a further aspect of the invention, a particular compound ofthe invention isN,N-dimethyl-2-morpholino-4-oxo-8-((1R)-1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;or a pharmaceutically-acceptable salt thereof.

According to a further aspect of the invention, a particular compound ofthe invention isN,N-dimethyl-2-morpholino-4-oxo-8-((1S)-1-(2,3,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide;or a pharmaceutically-acceptable salt thereof.

Another aspect of the present invention provides a process for preparinga compound of the Formula I, or a pharmaceutically-acceptable saltthereof. A suitable process is illustrated by the followingrepresentative process variants in which, unless otherwise stated, R¹,R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, n and R⁹ have any of the meanings definedhereinbefore. Necessary starting materials may be obtained by standardprocedures of organic chemistry. The preparation of such startingmaterials is described in conjunction with the following representativeprocess variants and within the accompanying Examples. Alternatively,necessary starting materials are obtainable by analogous procedures tothose illustrated which are within the ordinary skill of an organicchemist.

Suitable process variants include, for example, the following:—

(a) The reaction, conveniently in the presence of a suitable activatingreagent, of a compound of the Formula II

wherein R¹, R², n and R⁹ have any of the meanings defined hereinbeforeexcept that any functional group present is protected if necessary, withan amine derivative of the Formula III:

wherein R³ is H and R⁴, R⁵, R⁶, R⁷ and R⁸ have any of the meaningsdefined hereinbefore except that any functional group is protected ifnecessary, in the presence of a suitable base, to provide anintermediate compound, of the Formula IV:

which is then reduced by a suitable reducing agent, to form a compoundof the Formula I, whereafter any protecting group that is present isremoved.

A suitable activating reagent for the reaction includes, for example, aLewis acid such as tin (IV) tetrachloride, aluminium (III) trichlorideor titanium(IV) tetrachloride. Conveniently, the suitable catalyst istitanium(IV) tetrachloride.

Conveniently, the reaction is conducted in the presence of a suitablebase such as an organic base such as pyridine, 4-dimethylaminopyridine,triethylamine or N-methyl morpholine. Conveniently, the suitable base istriethylamine.

The reaction is conveniently carried out in the presence of a suitablesolvent or diluent such as for example, N,N-dimethylformamide,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene,xylene, halogenated solvents such as dichloromethane, chloroform orcarbon tetrachloride and at a temperature in the range, for example −50°C. to 100° C., preferably in the range 0° C. to 30° C.

A suitable reducing agent is a metal borohydride such as for examplesodium cyanotrihydroborate. The reduction reaction is convenientlycarried out in the presence of a suitable solvent or diluent, such asfor example an alcohol such as methanol or ethanol or a mixture ofsolvents containing alcohols, generally in the presence of a weak acidsuch as acetic acid. The reaction is conveniently carried out at atemperature in the range, for example, 0° C. to 30° C.

Compounds of the Formula II may, for example, be prepared by a crosscoupling reaction of a compound of the Formula V:

wherein n and R⁹ has any of the meanings defined hereinbefore exceptthat any functional group present is protected if necessary, with anamine compound of the Formula VI:

wherein R¹ and R² has any of the meanings defined hereinbefore, in thepresence of a suitable coupling agent such as, for example, TSTU(2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate), whereafter any protecting group that is present isremoved.

The reaction is conveniently carried out in the presence of a suitablebase. A suitable base is, for example, an organic amine base such as,for example, pyridine, 2,6-lutidine, collidine, 4-dimethylaminopyridine,triethylamine, N-methylmorpholine, diazabicyclo[5.4.0]undec-7-ene,diisopropylethyl amine, or, for example, an alkali or alkaline earthmetal carbonate, for example sodium carbonate, potassium carbonate orcalcium carbonate.

The reaction is conveniently carried out in the presence of a suitableinert solvent or diluent such as for example, N,N-dimethylformamide,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene,xylene, methanol ethanol, halogenated solvents such as dichloromethane,chloroform or carbon tetrachloride and at a temperature in the range,for example −50° C. to 100° C., preferably in the range 0° C. to 30° C.

Compounds of the Formula III may be obtained by conventional proceduresor are commercially available, known in the literature, or they can beprepared by standard processes known in the art.

Compounds of the Formula V may be obtained by analogous procedures tothose described in Example 1.00 herein, where the method for preparingthe starting material8-acetyl-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamideis given. In particular, compounds of the Formulae V may be obtained byprocedures in accordance with the following scheme:

For example, compounds of the Formula V may, be prepared by reaction ofa compound of the Formula VII:

wherein n and R⁹ has any of the meanings defined hereinbefore exceptthat any functional group present is protected if necessary, with asuitable stannane such as, for example, tributyl(1-ethoxyvinyl)stannaneunder Stille type conditions (for further details of such conditions seefor example: ‘Metal-Catalyzed Cross-Coupling Reactions’, Second Edition,Edited by Armin Meijere, Frangois Diederich, Wiley-VCH, 2004, Volume 1,p125), whereafter any protecting group that is present is removed.

A suitable catalyst for the reaction includes, for example, a metalliccatalyst such as a palladium(0), palladium(II) for exampletetrakis(triphenylphosphine)palladium(0), palladium(II) chloride,palladium(II) bromide, bis(triphenylphosphine)palladium(II) chloride,[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II),tris(dibenzilideneacetone)dipalladium. Optionally, the catalyst can beformed in-situ by the reaction of one or more of the above catalystswith a trialkylphosphine, such as, for example, tri-N-butylphosphine ortricyclohexylphosphine.

The reaction is conveniently carried out in the presence of a suitablesolvent or diluent such as for example, N,N-dimethylformamide,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene,xylene and at a temperature in the range, for example 20° C. to 150° C.,preferably in the range 60° C. to 120° C.

Compounds of the Formula VII may, for example, be prepared by reactionof a compound of the Formula VIII:

wherein R¹⁰ is (1-6C)alkyl, conveniently methyl or ethyl, with acompound of the Formula IX:

wherein n and R⁹ have any of the meanings defined hereinbefore, in thepresence of a suitable activating agent such as, for example, a Lewisacid, such as for example boron trifluoride-diethyl etherate, to providea compound, of the Formula VIIIa:

whereafter a saponifcation reaction can be used to form the compound ofthe Formula VII.

Reaction of the compounds of the Formula VIII with those of the FormulaIX is conveniently carried out in the presence of a suitable solvent ordiluent such as for example, N,N-dimethylformamide, tetrahydrofuran,1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene, xylene, halogenatedsolvents such as dichloromethane, chloroform or carbon tetrachloride andat a temperature in the range, for example 20° C. to 150° C., preferablyin the range 60° C. to 120° C.

The saponification reaction can be conducted for example by treatmentwith an alkali or alkaline earth metal hydroxide such as lithium,potassium or sodium hydroxide in a suitable solvent such as for example,methanol or a mixture of ethanol and water or a water miscible solvent,such as for example tetrahydrofuran or dioxane, at a temperature in therange, for example 0° C. to −100° C., preferably in the range 20-40° C.

Compounds of the Formula VIII have been described in the literature(Ger. Offen, DE 4318756, 1994 and Aust. J. Chem. 2003, 56, 1099), orthey can be prepared by standard processes known in the art.

Compounds of the Formula VIIIa may alternatively be obtained byprocedures in accordance with the following scheme, which has beendescribed in more detail in Example 1.00 herein, where the method forpreparing methyl 8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylateusing such a method is provided:

where, DCM is dichloromethane, LiHMDS is Lithium bis(trimethylsilyl)amide, EtOH is ethanol, Dppf is 1,1′-bis(diphenylphosphino)ferrocene, TEA is triethylamine, THF istetrahydrofuran and Tf₂O is Trifluoromethanesulfonic anhydride.

For example, compounds of the Formula VIIIa may, be prepared by reactionof a compound of the Formula VIII:

wherein R¹⁰ is (1-6C)alkyl, conveniently methyl or ethyl, with acompound of the Formula IXa:

wherein n and R⁹ have any of the meanings defined hereinbefore, in thepresence of a suitable activating agent such as, for example, a strongbase, such as for example Lithium bis(trimethyl silyl)amide, to providea compound, of the Formulae VIIIb:

whereafter a ring-closing reaction can be performed to form the compoundof the Formula VIIIa.

Reaction of the compounds of the Formula VIII with those of the FormulaIXa is conveniently carried out in the presence of a suitable solvent ordiluent such as for example, tetrahydrofuran, 1,4-dioxane,1,2-dimethoxyethane, benzene, toluene or xylene and at a temperature inthe range, for example −100° C. to ambient temperature, preferably inthe range −80° C. to 20° C.

The ring-closing reaction to convert a compound of the Formula VIIIbinto a compound of the Formula VIIIa can be conducted for example bytreatment with a dehydrating agent, such as for exampletrifluoromethanesulfonic anhydride, in a suitable solvent such as forexample dichloroethane at a temperature in the range, for example 0° C.to −100° C., conveniently in the range 20-60° C.

Alternatively, the Compound of the Formula VIIIa may be prepared inaccordance with the following scheme, which has been described in moredetail in Example 9.0 herein, which describes the preparation of methyl8-bromo-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate:

Alternatively, Compounds of the Formula II may, for example, be preparedby reaction of a compound of the Formula XVI:

wherein R¹, R², n and R⁹ have any of the meanings defined hereinbeforeexcept that any functional group present is protected if necessary, witha suitable stannane such as, for example,tributyl(1-ethoxyvinyl)stannane under Stille type conditions (forfurther details of such conditions see for example: ‘Metal-CatalyzedCross-Coupling Reactions’, Second Edition, Edited by Armin Meijere,Frangois Diederich, Wiley-VCH, 2004, Volume 1, p125), whereafter anyprotecting group that is present is removed.

A suitable catalyst for the Stille reaction includes, for example, ametallic catalyst such as a palladium(0), palladium(II) for exampletetrakis(triphenylphosphine)palladium(0), palladium(II) chloride,palladium(II) bromide, bis(triphenylphosphine)palladium(II) chloride,[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II),tris(dibenzilideneacetone)dipalladium. Optionally, the catalyst can beformed in-situ by the reaction of one or more of the above catalystswith a trialkylphosphine, such as, for example, tri-N-butylphosphine ortricyclohexylphosphine.

The reaction is conveniently carried out in the presence of a suitablesolvent or diluent such as for example, N,N-dimethylformamide,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene,xylene and at a temperature in the range, for example 20° C. to 150° C.,conveniently in the range 60° C. to 120° C.

Alternatively, Compounds of the Formula II may, for example, be preparedby reaction of a compound of the Formula XVI:

wherein R¹, R², n and R⁹ have any of the meanings defined hereinbeforeexcept that any functional group present is protected if necessary, witha suitable alkene such as for example, (1-vinyloxy)butane under Hecktype conditions whereafter any protecting group that is present isremoved.

A suitable catalyst for the Heck reaction includes, for example, ametallic catalyst such as a palladium(0), palladium(II) for exampletetrakis(triphenylphosphine)palladium(O), palladium(II) chloride,palladium(II) bromide, bis(triphenylphosphine)palladium(II) chloride,[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II),tris(dibenzilideneacetone)dipalladium; Optionally, the catalyst can beformed in-situ by the reaction of one or more of the above catalystswith a trialkylphosphine, such as, for example, tri-N-butylphosphine ortricyclohexylphosphine.

Conveniently, the catalyst is palladium (II) acetate in the presence ofbis (1,3-diphenylphosphino)propane.

The reaction is conveniently carried out in the presence of a suitablesolvent or diluent such as for example, N,N-dimethylformamide,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene,xylene or alcohols and at a temperature in the range, for example 20° C.to 150° C. Conveniently ethyleneglycol is used and the reaction isconducted at a temperature of between 90-130° C.

An example of a process scheme that may be used for the synthesis of acompound of the Formula XVI, such as for example8-bromo-N,N-dimethyl-2-(morpholin-4-yl)-4-oxo-4H-chromene-6-carboxamide,is the following:

where, DCM is dichloromethane, LiHMDS is Lithium bis(trimethylsilyl)amide, EtOH is ethanol, DIPEA is diisopropylethylamine, THF istetrahydrofuran, Tf₂O is Trifluoromethanesulfonic anhydride and TBTU is2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate.(b) The cross coupling reaction, conveniently in the presence of asuitable catalyst as defined hereinbefore, of a compound of the FormulaX:

wherein R³, R⁴, R⁵, R⁶, R⁷ and R⁸, n and R⁹ have any of the meaningsdefined hereinbefore except that any functional group present isprotected if necessary, conveniently in the presence of a suitable base,with an amine of the Formula VI:

wherein R¹ and R² have any of the meanings defined hereinbefore exceptthat any functional group is protected if necessary, in the presence ofa suitable coupling agent such as, for example, TSTU(2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate) or TBTU(2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate), whereafter any protecting group that is present isremoved.

The reaction is conveniently carried out in the presence of a suitablebase. A suitable base is, for example, an organic amine base such as,for example, pyridine, 2,6-lutidine, collidine, 4-dimethylaminopyridine,triethylamine, N-methylmorpholine, diazabicyclo[5.4.0]undec-7-ene,diisopropylethyl amine, or, for example, an alkali or alkaline earthmetal carbonate or hydroxide, for example sodium carbonate, potassiumcarbonate, calcium carbonate, sodium hydroxide or potassium hydroxide.

The reaction is conveniently carried out in the presence of a suitableinert solvent or diluent such as for example, N,N-dimethylformamide,N-methylpyrrolidone, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane,benzene, toluene, xylene, methanol ethanol, halogenated solvents such asdichloromethane, chloroform or carbon tetrachloride and at a temperaturein the range, for example −50° C. to 100° C., preferably in the range 0°C. 20 to 30° C.

Compounds of the Formula X may, for example, be prepared by asaponification reaction, of a compound of the Formula Xa:

wherein R³, R⁴, R⁵, R⁶, R⁷, R⁸ n and R⁹ has any of the meanings definedhereinbefore and R¹⁰ is (1-6C)alkyl, conveniently methyl or ethyl.

The saponification reaction can be conducted for example by treatment ofa compound of Formula Xa with an alkali or alkaline earth metalhydroxide such as lithium, potassium or sodium hydroxide in a suitablesolvent such as for example a mixture of ethanol and water or a watermiscible solvent, such as for example tetrahydrofuran or dioxane, at atemperature in the range, for example 0° C. to −100° C., preferably inthe range 20-40° C.

Compounds of the Formula Xa may, for example, be prepared by thereaction, conveniently in the presence of a suitable catalyst as definedhereinbefore, of a compound of the Formula XI:

wherein n and R⁹ has any of the meanings defined hereinbefore, R¹⁰ is(1-6C)alkyl, conveniently methyl or ethyl, and L is a displaceablegroup, with an amine derivative of the Formula III

wherein R³, R⁴, R⁵, R⁶, R⁷ and R⁸ has any of the meanings definedhereinbefore, whereafter any protecting group that is present isremoved.

A suitable displaceable group L is, for example, a halogeno group suchas a chloro, bromo, iodo group, trifluoromethanesulphonyl ormethanesulphonyl. Conveniently, the displaceable group L is bromo.

The reaction is conveniently carried out in the presence of a suitableinert solvent or diluent such as for example, N,N-dimethylformamide,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene,xylene, methanol ethanol, halogenated solvents such as dichloromethane,chloroform or carbon tetrachloride and at a temperature in the range,for example −50° C. to 100° C., preferably in the range 0° C. to 30° C.

Compounds of the Formulae XI may be obtained by analogous procedures tothose described in the Example 2.00 herein, where the method forpreparing the starting material8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid is given. In particular, compounds of the Formulae XI may beobtained by procedures in accordance with the following scheme:

For example, compounds of the Formula XI may, be prepared by reaction ofa compound of the Formula XII:

wherein n and R⁹ has any of the meanings and R¹⁰ is (1-6C)alkyl,conveniently methyl or ethyl, with an agent such as a halogenatingagent, for example a brominating agent, such as for example phosphorustribromide.

The reaction is conveniently carried out in the presence of a suitableinert solvent or diluent such as for example, N,N-dimethylformamide,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene,xylene, halogenated solvents such as DCM, chloroform or carbontetrachloride and at a temperature in the range, for example −50° C. to100° C., preferably in the range 0° C. to 30° C.

Compounds of the Formula XII may, for example, be prepared by reactionof a compound of the Formula XIII:

wherein n and R⁹ has any of the meanings and R¹⁰ is (1-6C)alkyl,conveniently methyl or ethyl, with a reducing agent such as a hydride,for example a borohydride derived reagent, such as sodium borohydride.

The reaction is conveniently carried out in the presence of a suitablesolvent or diluent such as for example alcohols (methanol, ethanol) or amixture of solvents containing alcohols and at a temperature in therange, for example −50° C. to 50° C., preferably in the range 0° C. to20° C.

Compounds of the Formula XIII may be obtained by coupling compounds ofthe Formula VIIIa with a suitable stannane such as, for example,tributyl(1-ethoxyvinyl)stannane under Stille type conditions usingconditions reported in process variant (a). Alternatively, Compounds ofthe Formula XIII may be obtained by coupling compounds of the FormulaVIIIa with a suitable alkene such as for example, (1-vinyloxy)butaneunder Heck type conditions (see conditions reported in process variant(a)).

(c) The reaction, conveniently in the presence of a suitable catalyst asdefined hereinbefore in process variant (b) above, of a compound of theFormula XIV:

wherein R¹, R², n and R⁹ have any of the meanings defined hereinbeforeexcept that any functional group present is protected if necessary and Lis a displaceable group, with an amine derivative of the Formula III

wherein R³, R⁴, R⁵, R⁶, R⁷ and R⁸ has any of the meanings definedhereinbefore, whereafter any protecting group that is present isremoved.

A suitable displaceable group L is, for example, a halogeno group suchas a chloro, bromo, iodo group, a methanesulphonyl or atrifluoromethanesulphonyl group. Conveniently, the displaceable group Lis bromo.

The reaction is conveniently carried out in the presence of a suitableinert solvent or diluent such as for example, N,N-dimethylformamide,N-methylpyrrolidone, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane,benzene, toluene, xylene, methanol ethanol, halogenated solvents such asdichloromethane, chloroform or carbon tetrachloride and at a temperaturein the range, for example −50° C. to 100° C., preferably in the range 0°C. to 50° C. Conveniently, the suitable inert solvent isN,N-dimethylformamide.

Compounds of the Formula XIV may, for example, be prepared by thereaction, conveniently in the presence of a suitable catalyst as definedhereinbefore, of a compound of the Formula XV:

wherein R¹, R², n and R⁹ has any of the meanings defined hereinbefore,with an agent such as a halogenating agent, for example a brominatingagent, such as for example phosphorus tribromide. The reaction isconveniently carried out in the presence of a suitable inert solvent ordiluent such as for example, N,N-dimethylformamide, tetrahydrofuran,1,4-dioxane, 1,2-dimethoxyethane, benzene, toluene, xylene, halogenatedsolvents such as dichloromethane, chloroform or carbon tetrachloride andat a temperature in the range, for example −50° C. to 100° C.,preferably in the range 0° C. to 30° C.

Compounds of the Formula XV may, for example, be prepared by a crosscoupling reaction, conveniently in the presence of a suitable catalystas defined hereinbefore in process variant (b) above, of a compound ofthe Formula XVa:

wherein n and R⁹ have any of the meanings defined hereinbefore exceptthat any functional group present is protected if necessary,conveniently in the presence of a suitable base, with an amine of theFormula VI:

wherein R¹ and R² have any of the meanings defined hereinbefore exceptthat any functional group is protected if necessary, in the presence ofa suitable coupling agent such as, for example, TSTU(2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate) or TBTU(2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate), whereafter any protecting group that is present isremoved.

The reaction is conveniently carried out in the presence of a suitablebase. A suitable base is, for example, an organic amine base such as,for example, pyridine, 2,6-lutidine, collidine, 4-dimethylaminopyridine,triethylamine, N-methylmorpholine, diazabicyclo[5.4.0]undec-7-ene,diisopropylethyl amine, or, for example, an alkali or alkaline earthmetal carbonate or hydroxide, for example sodium carbonate, potassiumcarbonate, calcium carbonate, sodium hydroxide or potassium hydroxide.

The reaction is conveniently carried out in the presence of a suitableinert solvent or diluent such as for example, N,N-dimethylformamide,N-methylpyrrolidone, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane,benzene, toluene, xylene, methanol ethanol, halogenated solvents such asdichloromethane, chloroform or carbon tetrachloride and at a temperaturein the range, for example −50° C. to 100° C., preferably in the range 0°C. to 30° C.

Compounds of the Formula XVa may, for example, be prepared by asaponification reaction, of a compound of the Formula XII:

wherein n and R⁹ has any of the meanings and R¹⁰ is (1-6C)alkyl,conveniently methyl or ethyl.

The saponification reaction can be conducted for example by treatment ofa compound of Formula XII with an alkali or alkaline earth metalhydroxide such as lithium, potassium or sodium hydroxide in a suitablesolvent such as for example methanol, or a mixture of ethanol and wateror a water miscible solvent, such as for example tetrahydrofuran ordioxane, at a temperature in the range, for example 0° C. to −100° C.,preferably in the range 20-40° C.

Alternatively, compounds of the Formula XV may, for example, be prepareda reduction reaction of a compound of the Formula II, where a suitablereducing agent is employed, such as for example a metal borohydride suchas for example sodium borohydride. The reaction is conveniently carriedout in the presence of a suitable solvent or diluent, such as forexample an alcohol such as methanol or ethanol or a mixture of solventscontaining alcohols, generally in the presence of a weak acid such asacetic acid. The reaction is conveniently carried out at a temperaturein the range, for example, 0° C. to 30° C.

(d) The reaction of a compound of the Formula XVII:

wherein R¹, R², R³, n and R⁹ have any of the meanings definedhereinbefore except that any functional group present is protected, withcompound of the Formula IIIa

wherein R⁴, R⁵, R⁶, R⁷ and R⁸ has any of the meanings definedhereinbefore and LG is a suitable leaving group, such as for example, ahalogeno group such as a chloro, bromo, iodo group (conveniently bromoor iodo), whereafter any protecting group that is present is removed.

Suitable reactions of this type are described as palladium type couplingBuchwald reactions in ‘Metal-Catalyzed Cross-Coupling Reactions’, SecondEdition, Edited by Armin Meijere, Francois Diederich, Wiley-VCH, 2004,Volume 1, p699).

Alternatively, compounds of the Formula I can be prepared by Chan-Lamcoupling type reactions, in which a compound of the Formula XVII isreacted with a compound of the Formula IIIb:

wherein, R¹¹ is (1-3C)alkyl or H. Such a reaction is convenientlycatalysed by a copper source, such as for example copper (II) acetate inDCM and is carried out by way of exposure to atmospheric oxygen atambient temperature (Tetrahedron Letters, 1998, 2933).

Compounds of the Formula XVII may, for example, be prepared by areductive amination reaction of a compound of the Formula II:

wherein R¹, R², n and R⁹ have any of the meanings defined hereinbeforeexcept that any functional group present is protected if necessary, withan amine of the Formula R₃NH₂ or an equivalent, wherein R³ has any ofthe meanings defined hereinbefore, in the presence of a reducing agent,whereafter any protecting group that is present is removed.

Where the compound of the Formula I is a single optically activeenantiomer (chiral centre being the carbon atom to which the groupsmethyl and —N-phenyl(R⁴)(R⁵)(R⁶)(R⁷)(R⁸) are attached), analogousprocedures to those outlined in the following scheme could be utilisedto prepare compounds of the Formula XVII (for more details of aparticular example of this, see Example 8.00 herein):

where, Ti(OEt)₄ is titanium (IV) tetraethoxide, (R)—H2NSOtBu is(R)-2-methylpropane-2-sulfinamide, DCM is dichloromethane, MeOH ismethanol and THF is tetrahydrofuran.

For example, a single optically active enantiomer of the Formula XVIImay, be prepared by a reaction of a compound of the Formula II with achiral equivalent of R₃NH₂ such as an chiral sulfinamide for example,(R)-2-methylpropane-2-sulfinamide, to give a corresponding imine. Thisreaction is conveniently carried out in the presence of a Lewis acidsuch as titanium (IV) tetraethoxide The reaction is conveniently carriedout in the presence of a suitable inert solvent or diluent such as forexample, N,N-dimethylformamide, tetrahydrofuran, 1,4-dioxane,1,2-dimethoxyethane, benzene, toluene, xylene, halogenated solvents suchas dichloromethane, chloroform or carbon tetrachloride and at atemperature in the range, for example −50° C. to 100° C., preferably inthe range 0° C. to 30° C.

Reduction of the corresponding imine is then performed using a reducingagent such as a borohydride, for example, sodium cyanoborohydride,whereafter any protecting group that is present is removed. The reactionis conveniently carried out in the presence of a weak acid. A suitableweak acid is, for example, acetic acid. The reaction is convenientlycarried out in the presence of an alcohol such as methanol or ethanol,or a mixture of an alcohol and an suitable inert solvent or diluent suchas for example, N,N-dimethylformamide, tetrahydrofuran, 1,4-dioxane,1,2-dimethoxyethane, benzene, toluene, xylene, halogenated solvents suchas dichloromethane, chloroform or carbon tetrachloride and at atemperature in the range, for example −50° C. to 20° C., preferablyaround −15° C.

It is to be understood that other permutations of the process steps inthe process variants described above are also possible. For example, aCompound of Formula I could be prepared using analogous procedures tothose described in process variants (a) to (d), but wherein the finalstep in the procedure is the introduction of the morpholine-(R⁹)_(n)group.

It is to be understood that any compound of Formula I obtained by any ofthe processes described hereinbefore can be converted into anothercompound of the Formula I if required. For example, a compound of theFormula I wherein R³ is H can be converted into a compound of theFormula I in which R³ is (1-3C)alkyl by alkylation with an alkylatingagent. For example, where R³ is methyl, a suitable alkylating agent suchas dimethyl sulphate or methyl iodide could be used. The reaction can beperformed in the presence of a strong base, such as for example sodiumbis(trimethylsikyl)amide optionally in the presence of a suitablecrown-ether (for example 1,4,7,10,13-pentaoxacyclopentadecane, alsoknown as 15-crown-5 for sodium) at low temperature (−78° C. to 0° C.) inan inert solvent such as for example THF.

When a pharmaceutically-acceptable salt of a chromenone derivative ofthe Formula I is required, for example an acid-addition salt, it may beobtained by, for example, reaction of said chromenone derivative with asuitable acid.

When a pharmaceutically-acceptable pro-drug of a chromenone derivativeof the Formula I is required, it may be obtained using a conventionalprocedure. For example, an in vivo cleavable ester of a chromenonederivative of the Formula I may be obtained by, for example, reaction ofa compound of the Formula I containing a carboxy group with apharmaceutically-acceptable alcohol or by reaction of a compound of theFormula I containing a hydroxy group with a pharmaceutically-acceptablecarboxylic acid. For example, an in vivo cleavable amide of a chromenonederivative of the Formula I may be obtained by, for example, reaction ofa compound of the Formula I containing a carboxy group with apharmaceutically-acceptable amine or by reaction of a compound of theFormula I containing an amino group with a pharmaceutically-acceptablecarboxylic acid.

It will also be appreciated by the person skilled in the organicsynthetic arts that certain of the ring substituents in the compounds ofthe present invention may be introduced by standard aromaticsubstitution reactions or generated by conventional functional groupmodifications either prior to or immediately following the processesmentioned above, and as such are included in the process aspect of theinvention. Such reactions and modifications include, for example,introduction of a substituent by means of an aromatic substitutionreaction, reduction of substituents, alkylation of substituents,acylation of substituents, amidation of substituents and oxidation ofsubstituents. The reagents and reaction conditions for such proceduresare well known in the chemical art. Particular examples of aromaticsubstitution reactions include the introduction of a nitro group usingconcentrated nitric acid, the introduction of an acyl group using, forexample, an acyl halide and Lewis acid (such as aluminium trichloride)under Friedel Crafts conditions; the introduction of an alkyl groupusing an alkyl halide and Lewis acid (such as aluminium trichloride)under Friedel Crafts conditions; and the introduction of a halogenogroup. Particular examples of modifications include the reduction of anitro group to an amino group by for example, catalytic hydrogenationwith a nickel catalyst or treatment with iron in the presence ofhydrochloric acid with heating; oxidation of alkylthio to alkylsulphinylor alkylsulphonyl.

It will also be appreciated that, in some of the reactions mentionedhereinbefore, it may be necessary or desirable to protect any sensitivegroups in the compounds. The instances where protection is necessary ordesirable and suitable methods for protection are known to those skilledin the art. Conventional protecting groups may be used in accordancewith standard practice (for illustration see T. W. Green, ProtectiveGroups in Organic Synthesis, John Wiley and Sons, 1991). Thus, ifreactants include groups such as amino, carboxy or hydroxy, it may bedesirable to protect the group in some of the reactions mentionedherein.

A suitable protecting group for an amino or alkylamino group is, forexample, an acyl group, for example an alkanoyl group such as acetyl, analkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl ort-butoxycarbonyl group, an arylmethoxycarbonyl group, for examplebenzyloxycarbonyl, or an aroyl group, for example benzoyl. Thedeprotection conditions for the above protecting groups necessarily varywith the choice of protecting group. Thus, for example, an acyl groupsuch as an alkanoyl or alkoxycarbonyl group or an aroyl group may beremoved for example, by hydrolysis with a suitable base such as analkali metal hydroxide, for example lithium or sodium hydroxide.Alternatively an acyl group such as a t-butoxycarbonyl group may beremoved, for example, by treatment with a suitable acid as hydrochloric,sulphuric or phosphoric acid or trifluoroacetic acid and anarylmethoxycarbonyl group such as a benzyloxycarbonyl group may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon, or by treatment with a Lewis acid for example borontris(trifluoroacetate). A suitable alternative protecting group for aprimary amino group is, for example, a phthaloyl group which may beremoved by treatment with an alkylamine, for exampledimethylaminopropylamine, or with hydrazine.

A suitable protecting group for a hydroxy group is, for example, an acylgroup, for example an alkanoyl group such as acetyl, an aroyl group, forexample benzoyl, or an arylmethyl group, for example benzyl. Thedeprotection conditions for the above protecting groups will necessarilyvary with the choice of protecting group. Thus, for example, an acylgroup such as an alkanoyl or an aroyl group may be removed, for example,by hydrolysis with a suitable base such as an alkali metal hydroxide,for example lithium or sodium hydroxide. Alternatively an arylmethylgroup such as a benzyl group may be removed, for example, byhydrogenation over a catalyst such as palladium-on-carbon.

A suitable protecting group for a carboxy group is, for example, anesterifying group, for example a methyl or an ethyl group which may beremoved, for example, by hydrolysis with a base such as sodiumhydroxide, or for example a t-butyl group which may be removed, forexample, by treatment with an acid, for example an organic acid such astrifluoroacetic acid, or for example a benzyl group which may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon.

The protecting groups may be removed at any convenient stage in thesynthesis using conventional techniques well known in the chemical art.

Certain of the intermediates (for example, compounds of the Formulae II,IV, V, VII, VIIIa, VIIIb, X, Xa, XI, XII, XIII, XIV, XV, XVa, XVI, XVII)defined herein are novel and these are provided as a further feature ofthe invention. For example, compounds of the Formula VIIIa (wherein n,R⁹ and R¹⁰ have any of the meanings defined hereinbefore) may be usefulas intermediates in the preparation of particular compounds of theinvention:

Furthermore, the following compound may be useful as an intermediate inthe preparation of particular compounds of the invention:

methyl 8-bromo-2-morpholin-4-yl-4-oxo-4H-chromene-6-carboxylate

Biological Assays

The following assays can be used to measure the effects of the compoundsof the present invention as inhibitors of PI3 kinase enzymes, asinhibitors in vitro of phospho AKT (ser473) in MAD-MB-468 cells, asinhibitors in vivo of phospho AKT (ser473) in Swiss athymic nu/nu mice,and as inhibitors in vivo of tumour growth in Swiss athymic nu/nu micetransplanted with the human prostatic adenocarcinoma cell line, PC3.

(a) In Vitro Enzyme Inhibition Assay

The inhibition of PI3Kβ, PI3Kα, PI3Kγ and PI3Kδ was evaluated in aKinase Glo based enzyme activity assay using human recombinant enzymes.The assay measures depletion of ATP after incubation with enzyme, PIP2and ATP plus compound. ATP at the end of the reaction is detected byaddition of Kinase Glo reagent, in this the Ultra Glo™ luciferase(Promega) uses the ATP as a substrate to catalyze the mono-oxygenationof luciferin and the generation of light. A direct relationship existsbetween the luminescence measured with the Kinase-Glo Plus Reagent andthe amount of ATP remaining in a completed kinase reaction andluminescence is inversely related to kinase activity. Twelve differentcompound concentrations were tested and raw data from the inhibition ofPI3Kb, PI3Kα, PI3Kγ and PI3Kδ was plotted versus inhibitorconcentration.

Method Details.

Compounds in 100% DMSO were added to assay plates by acousticdispensing. PI3Kβ was added in a Tris buffer (50 mM Tris pH7.4, 0.05%CHAPS, 2.1 mM DTT, and 10 mM MgCl₂) and allowed to preincubate withcompound for 20 minutes prior to addition of substrate solutioncontaining PIP2 and ATP. The enzyme reaction was stopped after 80minutes by the addition of Kinase Glo detection solution. Plates wereleft for 30 minutes at room temperature then read on a PherastarInstrument (Luminescence ATP 384 program) setting gain on max well. Thefinal concentration of DMSO, ATP and PIP2 in the assay were, 1%, 8 μM,and 80 μM respectively.

Data Analysis

IC₅₀ values were calculated using a log curve fitting to a non-linearregression package, fitting raw data to inhibitor concentration. TheIC₅₀ value is the concentration of test compound that inhibited 50% ofenzyme activity.

(b) Protocol for Detection of Phospho AKT (Ser473) in MAD-MB-468 Cells

MDA-MB-468 cells (human breast adenocarcinoma ATCC HTB 132) are seededinto Greiner 384 well black flat-bottomed plates by automated cellculture robot (Selec T). Cells can also be maintained manually andseeded into plates using multidrop or Wellmate. Cells seeded at 1500cell/well in 40 μl of DMEN containing 10% FCS and 1% glutamine. Cellplates are incubated for 18 hours in a 37° C. incubator.

Compounds are dosed onto cells using an Echo acoustic dispenser, whichdispenses nl quantities of compound or DMSO. Compounds are dosed in a 12point concentration range from 30 μM top dose, 28 compounds are dosed onone plate. There are 17 DMSO only positive control wells per plate, and16 negative control well which have been dosed with a concentration ofreference compound that will knockout the pAKT signal. Plates areincubated at 37° C. for 2 hours, cells are the fixed by the addition of10 μl of a 3.7% Formaldehyde solution in a fume cupboard using aWellmate.

After 30 minutes to allow for fixation, the fixative and media areremoved and the plates washed with Proclin PBS/A using Tecan PW384 platewasher in a fume cupboard. Wells are blocked and permeabilised with theaddition of 40 μl of PBS containing 0.5% Tween20 and 1% marvel using aWellmate and incubated for 60 minutes at room temperature.

Permeabilisation and blocking buffer removed using Tecan PW384 platewasher, then 20 μl primary antibody solution added using a Wellmate. Theprimary antibody solution is a 1:500 dilution of Rabbit anti-phospho AKTSer 473 (Cell signalling technologies catalogue number #3787) in PBS/Tcontaining 1% marvel (dried milk powder) and incubated overnight at 4°C.

Plates are washed using a Tecan PW384 plate washer three times withPhosphate Buffered Saline+0.05% (v/v) Polysorbate20 and Proclin300(Supelco). 20 μl of secondary antibody solution is then added to eachwell using a Wellmate and incubated for 1 hour at Room Temp. Thesecondary antibody solution is a 1:1000 dilution of Alexa Fluor 488anti-Rabbit (Molecular Probes cat no A11008) diluted in PhosphateBuffered Saline+0.05% (v/v) Polysorbate20 containing 1% marvel. Platesare washed three times as before then 20 μl PBS added to each well andplates sealed with black plate sealer.

The plates are read on an Acumen reader as soon as possible. Using thissystem IC₅₀ values can be generated and quality of plates determined bycontrol wells. Reference compounds are run each time to monitor assayperformance.

(c) Protocol for Detection of Phospho AKT (Ser473) in Swiss AthymicNu/Nu Mice

Swiss athymic nu/nu mice can be transplanted s.c with human prostaticadenocarcinoma cell line PC3 (ATCC CRL1435) to determine anti-tumouractivity of PI3 kinase inhibitors. On day 0, 1×10⁶ cells in 50%Matrigel™ (BD Biosciences #354234) are injected s.c. on the left flankof the animals. Animals are randomised into required group sizes(typically 5 per treatment group) when tumours reach a volume of˜400-600 mm³ and treatment commences. Tumours are taken at terminationand flash frozen in liquid nitrogen and stored at −80° C. untilanalysis.

1 ml of lysis buffer plus phosphatase inhibitors Sigma #P2850, Sigma#P5726 (diluted 1:100) and protease inhibitors Sigma #P8340 (diluted1:200) is added to each tumour in a Fastprep tube. The tumours arehomogenised for 1 minute on the Fastprep machine and then left on icefor 10 minutes. Samples are spun for 10 minutes at 13,000 rpm in achilled centrifuge. Cleared lysates are then taken into fresh tubes and5 μl used for a protein determination assay. All tumour samples arediluted to same concentration so that 15 μg is run per lane on a 4-15%NuPAGE Bis-Tris gels (Invitrogen) for 90 minutes at 140 Volts. Thesamples are randomised so that gel effects are minimised. After blottingonto Nitrocellulose membranes they are blocked for one hour thenincubated overnight with a 1:500 dilution of antibody to either totalAKT (CST #9272) or phospho AKT-ser 473 (CST #9271). Blots are washedthree times in PBST before incubation for one hour at room temperaturewith a 1:2,000 dilution of anti-rabbit secondary HRP-linked antibody(CST #7074). Block and antibody incubation buffer is 5% dried milkpowder in PBS with 0.05% Polysorbate. Blots are washed three times inPBS/T then visualised using Pierce West Dura ECL kit and theChemiGenius. Bands are quantified and a ratio of phospho to total signalis obtained for each sample. The controls are averaged and eachtreatment sample is normalised to the averaged control value.

(d) Protocol for Detection of Tumour Growth Inhibition in HumanProstatic Adenocarcinoma Cell Line PC3 Transplanted Swiss Athymic Nu/NuMice

Swiss athymic nu/nu mice can be transplanted s.c with the humanprostatic adenocarcinoma cell line PC3 (ATCC CRL1435) to determineanti-tumour activity of PI3 kinase inhibitors. On day 0, 1×10⁶ cells in50% Matrigel (BDM) are injected s.c. on the left flank of the animals.Animals are randomised into groups of 10-15 when tumours reach a volumeof ˜200-300 mm³ and treatment commences. Animals are dosed for 2-4 weeksby peroral, intravenous or intra-peritoneal routes with compound (andoptionally a cyp inhibitor such as 1-aminobenzotriazole) in a suitablevehicle at defined doses. Tumours are usually measured twice weekly bycaliper and volume of tumours calculated using elliptical formula(pi/6×width×length).

Although the pharmacological properties of the compounds of the FormulaI vary with structural change as expected, in general activity possessedby compounds of the Formula I may be demonstrated at the followingconcentrations or doses in one or more of the above tests (a) and (b):—

-   -   Test (a):—IC₅₀ versus PI3Kβ in the range, for example, 1 nM-25        μM;    -   Test (b):—IC₅₀ versus cellular phospho AKT (ser473) in        MAD-MB-468 cells, in the range, for example, 1 nM-25 μM;

Conveniently, particular compounds of the invention possess activity atthe following concentrations or doses in one or more of the above tests(a) and (b):—

-   -   Test (a):—IC₅₀ versus PI3Kβ in the range, for example, 1 nM-10        μM;    -   Test (b):—IC₅₀ versus cellular phospho AKT (ser473) in        MAD-MB-468 cells, in the range, for example, 1 nM-20 μM;

Conveniently, particular compounds of the invention possess activity atthe following concentrations or doses in one or more of the above tests(a), (b), (c) and (d):—

-   -   Test (a):—IC₅₀ versus PI3Kβ in the range, for example, 1 nM-10        μM;    -   Test (b):—IC₅₀ versus cellular phospho AKT (ser473) in        MAD-MB-468 cells, in the range, for example, 1 nM-20 μM;    -   Test (c):—>50% inhibition of in vivo phospho AKT (ser473) in the        range, for example, 1-200 mg/kg/day;    -   Test (d):—xenograft activity in the range, for example, 1-200        mg/kg/day.

For example, the chromenone compound disclosed as Example 1.00 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 2 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 9 nM.

For example, the chromenone compound disclosed as Example 2.05 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 5 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 17 nM.

For example, the chromenone compound disclosed as Example 3.03 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 9 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 35 (37) nM. The valuegiven in brackets is a mean IC₅₀ value calculated from a larger numberof replicates than were used to calculate the first value quoted for thegiven Example compound.

For example, the chromenone compound disclosed as Example 3.04 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 11 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 12 nM.

For example, the chromenone compound disclosed as Example 3.06 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 6 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 9 nM.

For example, the chromenone compound disclosed as Example 3.06apossesses activity in Test (a) with an IC₅₀ versus PI3Kβ ofapproximately 3.7 μM; and activity in Test (b) with an IC₅₀ versuscellular phospho AKT (ser473) in MAD-MB-468 cells of approximately 13.5μM.

For example, the chromenone compound disclosed as Example 3.06bpossesses activity in Test (a) with an IC₅₀ versus PI3Kβ ofapproximately 2 nM; and activity in Test (b) with an IC₅₀ versuscellular phospho AKT (ser473) in MAD-MB-468 cells of approximately 5 (3)nM. The value given in brackets is a mean IC₅₀ value calculated from alarger number of replicates than were used to calculate the first valuequoted for the given Example compound.

For example, the chromenone compound disclosed as Example 3.07 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 29 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 0.58 μM.

For example, the chromenone compound disclosed as Example 3.11 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 4 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 27 nM.

For example, the chromenone compound disclosed as Example 4.02 possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 4 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 1 (2) nM. The value givenin brackets is a mean IC₅₀ value calculated from a larger number ofreplicates than were used to calculate the first value quoted for thegiven Example compound.

For example, the chromenone compound disclosed as Example 3.13apossesses activity in Test (a) with an IC₅₀ versus PI3Kβ ofapproximately 4.5 μM; and activity in Test (b) with an IC₅₀ versuscellular phospho AKT (ser473) in MAD-MB-468 cells of approximately 4.3μM.

For example, the chromenone compound disclosed as Example 5.0a possessesactivity in Test (a) with an IC₅₀ versus PI3Kβ of approximately 5 nM;and activity in Test (b) with an IC₅₀ versus cellular phospho AKT(ser473) in MAD-MB-468 cells of approximately 15 nM.

For example, the chromenone compounds disclosed within the Examplespossess activity in Test (a) at the levels illustrated in Table A.

TABLE A Example number PI3Kβ inhibition, IC₅₀ (μM) 1.00 0.002 1.01 0.0011.02 0.002 2.00 0.009 2.01 0.007 2.02 0.007 2.03 0.006 2.04 0.004 2.050.005 2.06 0.006 2.07 0.005 2.08 0.007 3.00 0.007 3.01 0.006 3.02 0.0113.03 0.009 3.03a 0.649 3.03b 0.003 3.04 0.011 3.04a 2.249 3.04b 0.0033.05 0.006 3.06 0.006 3.06a 3.695 3.06b 0.002 3.07 0.029 3.08 0.011 3.090.004 3.10 0.004 3.11 0.004 3.12 0.007 (0.005)* 3.13 0.006 3.13a 4.5373.13b 0.004 3.14 0.004 3.15 0.01 3.16 0.005 3.17 0.006 3.18 0.01 3.190.005 3.20 0.005 3.21 0.114 3.22 0.004 3.23 0.013 3.24 0.006 3.25 0.2123.26 0.004 3.27 0.009 3.28 0.004 3.29 0.005 3.30 0.004 3.31 0.006 3.320.008 3.33 0.005 3.34 0.004 3.35 0.031 3.36 0.024 3.37 0.012 3.38 0.0173.39 0.021 3.40 0.043 3.41 0.014 4.01 0.004 4.02 0.004 4.03 0.004 4.040.004 4.05 0.008 4.06 0.004 4.07 0.004 4.08 0.005 4.09 0.003 4.10 0.0054.11 0.002 4.12 0.004 4.13 0.009 4.14 0.004 4.15 0.004 4.16 0.009 4.170.005 4.18 0.004 4.19 0.023 4.20 0.005 4.21 0.004 4.22 0.004 4.23 0.0034.24 0.004 4.25 0.005 4.26 0.004 5.0a 0.005 5.0b 1.236 5.01 0.008 5.020.007 5.03 0.007 6.0 0.005 7.0 0.011 7.0a 3.381 7.0b 0.005 7.01a 0.0087.01b 4.315 7.02 0.004 8.0 0.003 8.01 0.005 8.02 0.002 9.0 0.005 9.010.004 9.01a 17.900 9.01b 0.003 9.02 0.011 9.02a 22.671 9.02b 0.007 9.030.064 10.01 0.012 10.02 0.011 10.03 0.009 11.00 0.021 *this is a meanIC₅₀ value calculated from a larger number of replicates than were usedto calculate the first value quoted for the given Example compound.

According to a further aspect of the invention there is provided apharmaceutical composition, which comprises a chromenone derivative ofthe Formula I, or a pharmaceutically-acceptable salt thereof, as definedhereinbefore in association with a pharmaceutically-acceptable diluentor carrier.

The compositions of the invention may be in a form suitable for oral use(for example as tablets, lozenges, hard or soft capsules, aqueous oroily suspensions, emulsions, dispersible powders or granules, syrups orelixirs), for topical use (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), for administration byinhalation (for example as a finely divided powder or a liquid aerosol),for administration by insufflation (for example as a finely dividedpowder) or for parenteral administration (for example as a sterileaqueous or oily solution for intravenous, subcutaneous, intraperitonealor intramuscular dosing) or as a suppository for rectal dosing.

The compositions of the invention may be obtained by conventionalprocedures using conventional pharmaceutical excipients, well known inthe art. Thus, compositions intended for oral use may contain, forexample, one or more colouring, sweetening, flavouring and/orpreservative agents.

The amount of active ingredient that is combined with one or moreexcipients to produce a single dosage form will necessarily varydepending upon the host treated and the particular route ofadministration. For example, a formulation intended for oraladministration to humans will generally contain, for example, from 1 mgto 1 g of active agent (more suitably from 1 to 250 mg, for example from1 to 100 mg) compounded with an appropriate and convenient amount ofexcipients which may vary from about 5 to about 98 percent by weight ofthe total composition.

The size of the dose for therapeutic or prophylactic purposes of acompound of the Formula I will naturally vary according to the natureand severity of the disease state, the age and sex of the animal orpatient and the route of administration, according to well knownprinciples of medicine.

In using a compound of the Formula I for therapeutic or prophylacticpurposes it will generally be administered so that a daily dose in therange, for example, 1 mg/kg to 100 mg/kg body weight is received, givenif required in divided doses. In general, lower doses will beadministered when a parenteral route is employed. Thus, for example, forintravenous administration, a dose in the range, for example, 1 mg/kg to25 mg/kg body weight will generally be used. Similarly, foradministration by inhalation, a dose in the range, for example, 1 mg/kgto 25 mg/kg body weight will be used. Oral administration is howeverpreferred, particularly in tablet form. Typically, unit dosage formswill contain about 10 mg to 0.5 g of a compound of this invention.

As stated above, it is known that PI 3-kinase enzymes contribute totumourigenesis by one or more of the effects of mediating proliferationof cancer and other cells, mediating angiogenic events and mediating themotility, migration and invasiveness of cancer cells. We have found thatthe chromenone derivatives of the present invention possess potentanti-tumour activity which it is believed is obtained by way ofinhibition of one or more of the Class I PI 3-kinase enzymes (such asthe Class Ia PI 3-kinase enzymes and/or the Class Ib PI 3-kinase enzyme)that are involved in the signal transduction steps which lead to theproliferation and survival of tumour cells and the invasiveness andmigratory ability of metastasising tumour cells.

Accordingly, the derivatives of the present invention are of value asanti-tumour agents, in particular as selective inhibitors of theproliferation, survival, motility, dissemination and invasiveness ofmammalian cancer cells leading to inhibition of tumour growth andsurvival and to inhibition of metastatic tumour growth. Particularly,the chromenone derivatives of the present invention are of value asanti-proliferative and anti-invasive agents in the containment and/ortreatment of solid tumour disease. Particularly, the compounds of thepresent invention are expected to be useful in the prevention ortreatment of those tumours which are sensitive to inhibition of one ormore of the multiple PI 3-kinase enzymes such as the Class Ia PI3-kinase enzymes and the Class Ib PI 3-kinase enzyme that are involvedin the signal transduction steps which lead to the proliferation andsurvival of tumour cells and the migratory ability and invasiveness ofmetastasising tumour cells. Further, the compounds of the presentinvention are expected to be useful in the prevention or treatment ofthose tumours which are mediated alone or in part by inhibition of PI3-kinase enzymes such as the Class Ia PI 3-kinase enzymes and the ClassIb PI 3-kinase enzyme, i.e. the compounds may be used to produce a PI3-kinase enzyme inhibitory effect in a warm blooded animal in need ofsuch treatment.

As stated hereinbefore, inhibitors of PI 3-kinase enzymes should be oftherapeutic value for treatment of, for example, cancer of the breast,colorectum, lung (including small cell lung cancer, non-small cell lungcancer and bronchioalveolar cancer) and prostate, and of cancer of thebile duct, bone, bladder, head and neck, kidney, liver, gastrointestinaltissue, oesophagus, ovary, pancreas, skin, testes, thyroid, uterus,cervix and vulva, and of leukaemias [including acute lymphocticleukaemia (ALL) and chronic myelogenous leukaemia (CML)], multiplemyeloma and lymphomas.

According to a further aspect of the invention there is provided achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore for use as a medicament in awarm-blooded animal such as man.

According to a further aspect of the invention, there is provided achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore for use in the production of ananti-proliferative effect in a warm-blooded animal such as man.

According to a further feature of this aspect of the invention there isprovided a chromenone derivative of the Formula I, or a pharmaceuticallyacceptable salt thereof, as defined hereinbefore for use in awarm-blooded animal such as man as an anti-invasive agent in thecontainment and/or treatment of solid tumour disease.

According to a further aspect of the invention, there is provided theuse of a chromenone derivative of the Formula I, or a pharmaceuticallyacceptable salt thereof, as defined hereinbefore for the production ofan anti-proliferative effect in a warm-blooded animal such as man.

According to a further feature of this aspect of the invention there isprovided the use of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in the production of ananti-proliferative effect in a warm-blooded animal such as man.

According to a further feature of this aspect of the invention there isprovided the use of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in a warm-blooded animal such as manas an anti-invasive agent in the containment and/or treatment of solidtumour disease.

According to a further feature of this aspect of the invention there isprovided a method for producing an anti-proliferative effect in a warmblooded animal, such as man, in need of such treatment which comprisesadministering to said animal an effective amount of a chromenonederivative of the Formula I, or a pharmaceutically acceptable saltthereof, as defined hereinbefore.

According to a further feature of this aspect of the invention there isprovided a method for producing an anti-invasive effect by thecontainment and/or treatment of solid tumour disease in a warm bloodedanimal, such as man, in need of such treatment which comprisesadministering to said animal an effective amount of a chromenonederivative of the Formula I, or a pharmaceutically acceptable saltthereof, solvate or pro-drug, as defined hereinbefore.

According to a further aspect of the invention, there is provided achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore for use in the prevention ortreatment of cancer in a warm blooded animal such as man.

According to a further aspect of the invention there is provided the useof a chromenone e derivative of the Formula I, or a pharmaceuticallyacceptable salt thereof, as defined hereinbefore in the manufacture of amedicament for use in the prevention or treatment of cancer in a warmblooded animal such as man.

According to a further feature of this aspect of the invention there isprovided a method for the prevention or treatment of cancer in a warmblooded animal, such as man, in need of such treatment which comprisesadministering to said animal an effective amount of a chromenonederivative of the Formula I, or a pharmaceutically acceptable saltthereof, as defined hereinbefore.

According to a further aspect of the invention there is provided the useof a chromenone e derivative of the Formula I, or a pharmaceuticallyacceptable salt thereof, as defined hereinbefore in the manufacture of amedicament for use in the prevention or treatment of solid tumourdisease in a warm blooded animal such as man.

According to a further feature of this aspect of the invention there isprovided a method for the prevention or treatment of solid tumourdisease in a warm blooded animal, such as man, in need of such treatmentwhich comprises administering to said animal an effective amount of achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore.

According to a further aspect of the invention there is provided achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore for use in the prevention ortreatment of those tumours which are sensitive to inhibition of PI3-kinase enzymes (such as the Class Ia enzymes and/or the Class Ib PI3-kinase enzyme) that are involved in the signal transduction stepswhich lead to the proliferation, survival, invasiveness and migratoryability of tumour cells.

According to a further feature of this aspect of the invention there isprovided the use of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in the prevention or treatment ofthose tumours which are sensitive to inhibition of PI 3-kinase enzymes(such as the Class Ia enzymes and/or the Class Ib PI 3-kinase enzyme)that are involved in the signal transduction steps which lead to theproliferation, survival, invasiveness and migratory ability of tumourcells.

According to a further feature of this aspect of the invention there isprovided a method for the prevention or treatment of those tumours whichare sensitive to inhibition of PI 3-kinase enzymes (such as the Class Iaenzymes and/or the Class Ib PI 3-kinase enzyme) that are involved in thesignal transduction steps which lead to the proliferation, survival,invasiveness and migratory ability of tumour cells which comprisesadministering to said animal an effective amount of a chromenonederivative of the Formula I, or a pharmaceutically acceptable saltthereof, as defined hereinbefore.

According to a further aspect of the invention there is provided achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore for use in providing a PI 3-kinaseenzyme inhibitory effect (such as a Class Ia PI 3-kinase enzyme or ClassIb PI 3-kinase enzyme inhibitory effect).

According to a further feature of this aspect of the invention there isprovided the use of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in providing a PI 3-kinase enzymeinhibitory effect (such as a Class Ia PI 3-kinase enzyme or Class Ib PI3-kinase enzyme inhibitory effect).

According to a further aspect of the invention there is also provided amethod for providing a PI 3-kinase enzyme inhibitory effect (such as aClass Ia PI 3-kinase enzyme or Class Ib PI 3-kinase enzyme inhibitoryeffect) which comprises administering an effective amount of achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore.

As stated hereinbefore, certain compounds of the present invention,possess substantially better potency against Class Ia PI 3-kinaseenzymes than against the Class Ib PI 3-kinase enzyme or against EGFreceptor tyrosine kinase, VEGF receptor tyrosine kinase or Srcnon-receptor tyrosine kinase enzymes. Such compounds possess sufficientpotency against Class Ia PI 3-kinase enzymes that they may be used in anamount sufficient to inhibit Class Ia PI 3-kinase enzymes whilstdemonstrating little activity against the Class Ib PI 3-kinase enzyme oragainst EGF receptor tyrosine kinase, VEGF receptor tyrosine kinase orSrc non-receptor tyrosine kinase enzymes. Such compounds are likely tobe useful for the selective inhibition of Class Ia PI 3-kinase enzymesand are likely to be useful for the effective treatment of, for exampleClass Ia PI 3-kinase enzyme driven tumours.

According to this aspect of the invention there is provided a chromenonederivative of the Formula I, or a pharmaceutically acceptable saltthereof, as defined hereinbefore for use in providing a selective ClassIa PI 3-kinase enzyme inhibitory effect.

According to a further feature of this aspect of the invention there isprovided the use of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in providing a selective Class Ia PI3-kinase enzyme inhibitory effect.

According to a further aspect of the invention there is also provided amethod for providing a selective Class Ia PI 3-kinase enzyme inhibitoryeffect which comprises administering an effective amount of a chromenonederivative of the Formula I, or a pharmaceutically acceptable saltthereof, as defined hereinbefore.

By “a selective Class Ia PI 3-kinase enzyme inhibitory effect” is meantthat the chromenone derivatives of the Formula I are more potent againstClass Ia PI 3-kinase enzymes than against other kinase enzymes. Inparticular, some of the compounds according to the invention are morepotent against Class Ia PI 3-kinase enzymes than against other kinasessuch as receptor or non-receptor tyrosine kinases or serine/threoninekinases. For example a selective Class Ia PI 3-kinase enzyme inhibitoraccording to the invention is at least 5 times more potent, convenientlyat least 10 times more potent, more conveniently at least 100 times morepotent, against Class Ia PI 3-kinase enzymes than against other kinases.

According to a further feature of the invention there is provided achromenone derivative of the Formula I, or a pharmaceutically acceptablesalt thereof, as defined hereinbefore for use in the treatment of cancerof the breast, colorectum, lung (including small cell lung cancer,non-small cell lung cancer and bronchioalveolar cancer) and prostate.

According to a further feature of this aspect of the invention there isprovided a chromenone derivative of the Formula I, or a pharmaceuticallyacceptable salt thereof, as defined hereinbefore for use in thetreatment of cancer of the bile duct, bone, bladder, head and neck,kidney, liver, gastrointestinal tissue, oesophagus, ovary, pancreas,skin, testes, thyroid, uterus, cervix and vulva, and of leukaemias(including ALL and CML), multiple myeloma and lymphomas.

According to a further feature of this aspect of the invention there isprovided the use of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in the treatment of cancer of thebreast, colorectum, lung (including small cell lung cancer, non-smallcell lung cancer and bronchioalveolar cancer) and prostate.

According to a further feature of this aspect of the invention there isprovided the use of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in the treatment of cancer of thebile duct, bone, bladder, head and neck, kidney, liver, gastrointestinaltissue, oesophagus, ovary, pancreas, skin, testes, thyroid, uterus,cervix and vulva, and of leukaemias (including ALL and CML), multiplemyeloma and lymphomas.

According to a further feature of this aspect of the invention there isprovided a method for treating cancer of the breast, colorectum, lung(including small cell lung cancer, non-small cell lung cancer andbronchioalveolar cancer) and prostate in a warm blooded animal such asman that is in need of such treatment which comprises administering aneffective amount of a chromenone derivative of the Formula I, or apharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further feature of this aspect of the invention there isprovided a method for treating cancer of the bile duct, bone, bladder,head and neck, kidney, liver, gastrointestinal tissue, oesophagus,ovary, pancreas, skin, testes, thyroid, uterus, cervix and vulva, and ofleukaemias (including ALL and CML), multiple myeloma and lymphomas in awarm blooded animal such as man that is in need of such treatment whichcomprises administering an effective amount of a chromenone derivativeof the Formula I, or a pharmaceutically acceptable salt thereof, asdefined hereinbefore.

As stated hereinbefore, the in vivo effects of a compound of the FormulaI may be exerted in part by one or more metabolites that are formedwithin the human or animal body after administration of a compound ofthe Formula I.

Particular compounds of the invention possess better potency against PI3-kinase β than against other class I PI 3-kinase isoforms such as α, γand δ.

The present invention therefore also contemplates a method forinhibiting phosphoinositide 3-kinase β in a patient, comprisingadministering to a patient an amount of the compound of formula (I), ora pharmaceutically acceptable salt thereof, effective in inhibiting thephosphoinositide 3-kinase β in the patient.

The compound of formula (I), or a pharmaceutically acceptable saltthereof, being an inhibitor of PI 3-kinase, also has potentialtherapeutic uses in a variety of other disease states. For example, PI3-kinase plays an important role in promoting smooth muscleproliferation in the vascular tree, i.e. vascular smooth muscle cells,Thyberg, 1998, European Journal of Cell Biology 76(1):33-42, and in thelungs (airway smooth muscle cells), Krymskaya, V. P., BioDrugs, 2007.21(2): 85-95. Excessive proliferation of vascular smooth muscle cellsplays an important role in the formation of atherosclerotic plaques andin the development of neointimal hyperplasia following invasive vascularprocedures, Scwartz et al., 1984, Progress in Cardiovascular Disease26:355-372; Clowes et al., 1978, Laboratory Investigations 39:141-150.Moreover, excessive proliferation of airway smooth muscle cells leads tothe development of COPD in the setting of asthma and chronic bronchitis.Inhibitors of PI 3-kinase activity therefore may be used to preventvascular restenosis, atherosclerosis, and COPD.

PI 3-kinases also play an important role in regulating tumor cells andin the propensity of these cells to undergo apoptosis growth (Sellers etal., 1999, The Journal of Clinical Investigation 104:1655-1661).Additionally, uncontrolled regulation of the PI 3-kinase lipid productsPI(3,4,5)P₃ and PI(3,4)P₂ by the lipid phosphatase PTEN plays animportant role in progression of a number of malignant tumors in humans(Leevers et al., 1999, Current Opinion in Cell Biology 11:219-225). Aspecific role for the phosphoinositide 3-kinase 0 (PI3KI3) isoform hasbeen described in these types of cancers (Jia S et al., 2008, Nature454(7205):₇₇₆₋₉; Wee et al., 2008, PNAS 105(35):₁₃₀₅₇₋₆₂). Therefore,the compound of formula (I), or a pharmaceutically acceptable saltthereof, being an inhibitor of PI 3-kinase, may be used to treatneoplasms in humans.

PI 3-kinase also plays an important role in leukocyte function (Fulleret al., 1999, The Journal of Immunology 162(11):6337-6340; Eder et al.,1998, The Journal of Biological Chemistry 273(43):28025-31) andlymphocyte function (Vicente-Manzanares et al., 1999, The Journal ofImmunology 163(7):4001-4012). For example, leukocyte adhesion toinflamed endothelium involves activation of endogenous leukocyteintegrins by a PI 3-kinase-dependent signaling process. Furthermore,oxidative burst (Nishioka et al., 1998, FEBS Letters 441(1):63-66 andCondliffe, A. M., et al., Blood, 2005. 106(4):1432-40) and cytoskeletalreorganization (Kirsch et al., 1999, Proceedings National Academy ofSciences USA 96(11):6211-6216) in neutrophils appears to involve PI3-kinase signaling. Neutrophil migration and directional movement arealso dependent on PI 3-kinase activity (Camps, M., et al., Nat Med,2005. 11(9): p. 936-43 and Sadhu, C., et al., J Immunol, 2003. 170(5):2647-54). Thus, inhibitors of PI 3-kinase may be useful in reducingleukocyte adhesion and activation at sites of inflammation and thereforemay be used to treat acute and/or chronic inflammatory disorders. PI3-kinase also plays an important role in lymphocyte proliferation andactivation, Fruman et al., 1999, Science 283 (5400): 393-397. Given theimportant role of lymphocytes in auto-immune diseases, an inhibitor ofPI 3-kinase activity may be used in the treatment of such disorders.

The anti-cancer treatment defined hereinbefore may be applied as a soletherapy or may involve, in addition to the compound of the invention,conventional surgery or radiotherapy or chemotherapy. Such chemotherapymay include one or more of the following categories of anti-tumouragents:—

-   (i) other antiproliferative/antineoplastic drugs and combinations    thereof, as used in medical oncology, such as alkylating agents (for    example cis-platin, oxaliplatin, carboplatin, cyclophosphamide,    nitrogen mustard, melphalan, chlorambucil, busulphan, temozolamide    and nitrosoureas); antimetabolites (for example gemcitabine and    antifolates such as fluoropyrimidines like 5-fluorouracil and    tegafur, raltitrexed, methotrexate, cytosine arabinoside, and    hydroxyurea); antitumour antibiotics (for example anthracyclines    like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin,    idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic    agents (for example vinca alkaloids like vincristine, vinblastine,    vindesine and vinorelbine and taxoids like taxol and taxotere and    polokinase inhibitors); and topoisomerase inhibitors (for example    epipodophyllotoxins like etoposide and teniposide, amsacrine,    topotecan and camptothecin);-   (ii) cytostatic agents such as antioestrogens (for example    tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and    iodoxyfene), antiandrogens (for example bicalutamide, flutamide,    nilutamide and cyproterone acetate), LHRH antagonists or LHRH    agonists (for example goserelin, leuprorelin and buserelin),    progestogens (for example megestrol acetate), aromatase inhibitors    (for example as anastrozole, letrozole, vorazole and exemestane) and    inhibitors of 5α-reductase such as finasteride;-   (iii) anti-invasion agents [for example c-Src kinase family    inhibitors like    4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran-4-yloxyquinazoline    (AZD0530; International Patent Application WO 01/94341),    N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazole-5-carboxamide    (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661) and    bosutinib (SKI-606), and metalloproteinase inhibitors like    marimastat, inhibitors of urokinase plasminogen activator receptor    function or antibodies to Heparanase]; (iv) inhibitors of growth    factor function: for example such inhibitors include growth factor    antibodies and growth factor receptor antibodies (for example the    anti-erbB2 antibody trastuzumab [Herceptin™], the anti-EGFR antibody    panitumumab, the anti-erbB1 antibody cetuximab [Erbitux, C225] and    any growth factor or growth factor receptor antibodies disclosed by    Stem et al. Critical reviews in oncology/haematology, 2005, Vol. 54,    pp 11-29); such inhibitors also include tyrosine kinase inhibitors,    for example inhibitors of the epidermal growth factor family (for    example EGFR family tyrosine kinase inhibitors such as    N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine    (gefitinib, ZD1839),    N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine    (erlotinib, OSI-774) and    6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine    (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib);    inhibitors of the hepatocyte growth factor family; inhibitors of the    insulin growth factor family; inhibitors of the platelet-derived    growth factor family such as imatinib and/or nilotinib (AMN107);    inhibitors of serine/threonine kinases (for example Ras/Raf    signalling inhibitors such as famesyl transferase inhibitors, for    example sorafenib (BAY 43-9006), tipifamib (R115777) and lonafarnib    (SCH66336)), inhibitors of cell signalling through MEK and/or AKT    kinases, c-kit inhibitors, abl kinase inhibitors, PI3 kinase    inhibitors, Plt3 kinase inhibitors, CSF-1R kinase inhibitors, IGF    receptor (insulin-like growth factor) kinase inhibitors; aurora    kinase inhibitors (for example AZD1152, PH739358, VX-680, MLN8054,    R763, MP235, MP529, VX-528 AND AX39459) and cyclin dependent kinase    inhibitors such as CDK2 and/or CDK4 inhibitors;-   (v) antiangiogenic agents such as those which inhibit the effects of    vascular endothelial growth factor, [for example the anti-vascular    endothelial cell growth factor antibody bevacizumab (Avastin™) and    for example, a VEGF receptor tyrosine kinase inhibitor such as    vandetanib (ZD6474), vatalanib (PTK787), sunitinib (SU11248),    axitinib (AG-013736), pazopanib (GW 786034) and    4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)quinazoline    (AZD2171; Example 240 within WO 00/47212), compounds such as those    disclosed in International Patent Applications WO97/22596, WO    97/30035, WO 97/32856 and WO 98/13354 and compounds that work by    other mechanisms (for example linomide, inhibitors of integrin αvβ3    function and angiostatin)];-   (vi) vascular damaging agents such as Combretastatin A4 and    compounds disclosed in International Patent Applications WO    99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 and WO    02/08213;-   (vii) an endothelin receptor antagonist, for example zibotentan    (ZD4054) or atrasentan;-   (viii) antisense therapies, for example those which are directed to    the targets listed above, such as ISIS 2503, an anti-ras antisense;-   (ix) gene therapy approaches, including for example approaches to    replace aberrant genes such as aberrant p53 or aberrant BRCA1 or    BRCA2, GDEPT (gene-directed enzyme pro-drug therapy) approaches such    as those using cytosine deaminase, thymidine kinase or a bacterial    nitroreductase enzyme and approaches to increase patient tolerance    to chemotherapy or radiotherapy such as multi-drug resistance gene    therapy; and-   (x) immunotherapy approaches, including for example ex-vivo and    in-vivo approaches to increase the immunogenicity of patient tumour    cells, such as transfection with cytokines such as interleukin 2,    interleukin 4 or granulocyte-macrophage colony stimulating factor,    approaches to decrease T-cell anergy, approaches using transfected    immune cells such as cytokine-transfected dendritic cells,    approaches using cytokine-transfected tumour cell lines and    approaches using anti-idiotypic antibodies.

According to this aspect of the invention there is provided acombination suitable for use in the treatment of cancer comprising acompound of formula I as defined hereinbefore or a pharmaceuticallyacceptable salt thereof and any one of the anti tumour agents listedunder (i)-(ix) above.

Therefore in a further aspect of the invention there is provided acompound of formula (I) or a pharmaceutically acceptable salt thereof incombination with an anti-tumour agent selected from one listed under(i)-(ix) herein above.

In a further aspect of the invention there is provided a combinationsuitable for use in the treatment of cancer comprising a compound offormula I as defined hereinbefore or a pharmaceutically acceptable saltthereof and any one of the anti tumour agents listed under (i) above.

In a further aspect of the invention there is provided a combinationsuitable for use in the treatment of cancer comprising a compound offormula I as defined hereinbefore or a pharmaceutically acceptable saltthereof and a taxoid, such as for example taxol or taxotere,conveniently taxotere.

Herein, where the term “combination” is used it is to be understood thatthis refers to simultaneous, separate or sequential administration. Inone aspect of the invention “combination” refers to simultaneousadministration. In another aspect of the invention “combination” refersto separate administration. In a further aspect of the invention“combination” refers to sequential administration. Where theadministration is sequential or separate, the delay in administering thesecond component should not be such as to lose the beneficial effect ofthe combination.

According to a further aspect of the invention there is provided apharmaceutical composition which comprises a compound of formula (I) ora pharmaceutically acceptable salt thereof in combination with ananti-tumour agent selected from one listed under (i)-(ix) herein above,in association with a pharmaceutically acceptable diluent or carrier.

According to a further aspect of the invention there is provided apharmaceutical composition which comprises a compound of formula (I) ora pharmaceutically acceptable salt thereof in combination with ananti-tumour agent selected from one listed under (i)-(ix) herein above,in association with a pharmaceutically acceptable diluent or carrier foruse in treating cancer.

According to another feature of the invention there is provided the useof a compound of the formula (I) or a pharmaceutically acceptable saltthereof in combination with an anti-tumour agent selected from onelisted under (i)-(ix) herein above, in the manufacture of a medicamentfor use in cancer in a warm-blooded animal, such as man.

Therefore in an additional feature of the invention, there is provided amethod of treating cancer in a warm-blooded animal, such as man, in needof such treatment which comprises administering to said animal aneffective amount of a compound of formula (I) or a pharmaceuticallyacceptable salt thereof in combination with an anti-tumour agentselected from one listed under (i)-(ix) herein above.

According to a further aspect of the present invention there is provideda kit comprising a compound of formula (I) or a pharmaceuticallyacceptable salt thereof in combination with an anti-tumour agentselected from one listed under (i)-(ix) herein above.

According to a further aspect of the present invention there is provideda kit comprising:

-   a) a compound of formula (I) or a pharmaceutically acceptable salt    thereof in a first unit dosage form;-   b) an anti-tumour agent selected from one listed under (i)-(ix)    herein above; in a second unit dosage form; and-   c) container means for containing said first and second dosage    forms.

Although the compounds of the Formula I are primarily of value astherapeutic agents for use in warm-blooded animals (including man), theyare also useful whenever it is required to inhibit the effects of ClassI PI 3-kinase enzyme, particularly a Class Ia PI 3-kinase enzymes and/orClass Ib PI 3-kinase enzyme, more particularly a Class Ia PI 3-kinaseenzymes, which includes PI 3-kinase β. Thus, they are useful aspharmacological standards for use in the development of new biologicaltests and in the search for new pharmacological agents.

The invention will now be illustrated in the following Examples inwhich, generally:

(i) operations were carried out at ambient temperature, i.e. in therange 17 to 25° C. and under an atmosphere of an inert gas such asnitrogen unless otherwise stated;

(ii) evaporations were carried out by rotary evaporation or utilisingGenevac equipment in vacuo and work-up procedures were carried out afterremoval of residual solids by filtration;

(iii) Flash chromatography purifications were performed on an automatedArmen Glider Flash: Spot II Ultimate (Armen Instrument, Saint-Ave,France) using prepacked Merck normal phase Si60 silica cartridges(granulometry: 15-40 or 40-63 m) obtained from Merck, Darmstad, Germany;

(iv) preparative chromatography was performed on a Waters instrument(600/2700 or 2525) fitted with a ZMD or ZQ ESCi mass spectrometers and aWaters X-Terra or a Waters X-Bridge or a Waters SunFire reverse-phasecolumn (C-18, 5 microns silica, 19 mm diameter, 100 mm length, flow rateof 40 mL/minute) using decreasingly polar mixtures of water (containing0.2% ammonium carbonate) and acetonitrile as eluent;

(v) yields, where present, are not necessarily the maximum attainable;

(vi) in general, the structures of end-products of the Formula I wereconfirmed by nuclear magnetic resonance (NMR) spectroscopy; NMR chemicalshift values were measured on the delta scale [proton magnetic resonancespectra were determined using a Bruker Avance 500 (500 MHz) instrument];measurements were taken at ambient temperature unless otherwisespecified; the following abbreviations have been used: s, singlet; d,doublet; t, triplet; q, quartet; m, multiplet; dd, doublet of doublets;ddd, doublet of doublet of doublet; dt, doublet of triplets; bs, broadsignal;

(vii) in general, end-products of the Formula I were also characterisedby mass spectroscopy following liquid chromatography (LCMS); LCMS wascarried out using an Waters Alliance HT (2790 & 2795) fitted with aWaters ZQ ESCi or ZMD ESCi mass spectrometer and an X Bridge 5 μm C-18column (2.1×50 mm) at a flow rate of 2.4 mL/min, using a solvent systemof 95% A+5% C to 95% B+5% C over 4 minutes, where A=water, B=methanol,C=1:1 methanol:water (containing 0.2% ammonium carbonate);

(viii) intermediates were not generally fully characterised and puritywas assessed by thin layer chromatographic, mass spectral, HPLC and/orNMR analysis;

(ix) X-ray powder diffraction spectra were determined (using a Bruker D4Analytical Instrument) by mounting a sample of the crystalline materialon a Bruker single silicon crystal (SSC) wafer mount and spreading outthe sample into a thin layer with the aid of a microscope slide. Thesample was spun at 30 revolutions per minute (to improve countingstatistics) and irradiated with X-rays generated by a copper long-finefocus tube operated at 40 kV and 40 mA with a wavelength of 1.5418angstroms. The collimated X-ray source was passed through an automaticvariable divergence slit set at V20 and the reflected radiation directedthrough a 5.89 mm antiscatter slit and a 9.55 mm detector slit. Thesample was exposed for 0.03 seconds per 0.00570° 2-theta increment(continuous scan mode) over the range 2 degrees to 40 degrees 2-theta intheta-theta mode. The running time was 3 minutes and 36 seconds. Theinstrument was equipped with a Position sensitive detector (Lynxeye).Control and data capture was by means of a Dell Optiplex 686 NT 4.0Workstation operating with Diffrac+ software. Persons skilled in the artof X-ray powder diffraction will realise that the relative intensity ofpeaks can be affected by, for example, grains above 30 microns in sizeand non-unitary aspect ratios that may affect analysis of samples. Theskilled person will also realise that the position of reflections can beaffected by the precise height at which the sample sits in thediffractometer and the zero calibration of the diffractometer. Thesurface planarity of the sample may also have a small effect. Hence thediffraction pattern data presented are not to be taken as absolutevalues;

(x) Differential Scanning Calorimetry was performed using a TAInstruments Q1000 DSC instrument. Typically less than 5 mg of materialcontained in a standard aluminium pan fitted with a lid was heated overthe temperature range 25° C. to 300° C. at a constant heating rate of10° C. per minute. A purge gas using nitrogen was used at a flow rate of50 mL per minute; and

(xi) the following abbreviations have been used:—

-   -   aq. aqueous    -   CDCl₃ deutero-chloroform    -   CHCl3 chloroform    -   DCM dichloromethane    -   DEA diethyl amine    -   DIPEA N-ethyl-N-isopropylpropan-2-amine    -   DMF N,N-dimethylformamide    -   DMSO dimethyl sulphoxide    -   DSC Differential Scanning Calorimetry    -   DTAD (E)-di-tert-butyl diazene-1,2-dicarboxylate    -   EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride    -   Ether diethyl ether    -   % ee % enantiomeric excess    -   HOPO 2-hydroxy-pyridine n-oxide    -   IPA isopropyl alcohol    -   MeCN acetonitrile    -   MeOH methanol    -   MTBE methyl tert-butyl ether    -   NMP 1-methyl-2-pyrrolidone    -   sat. saturated    -   sol. solution    -   THF tetrahydrofuran    -   TEA triethyl amine    -   TBTU        2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouronium        tetrafluoroborate    -   TSTU 2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouronium        tetrafluoroborate.

EXAMPLE 1.00N-(2-(dimethylamino)ethyl)-8-(1-(4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

Titanium(IV) chloride (0.129 mL, 0.13 mmol) was added to a stirredsuspension of8-acetyl-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(100 mg, 0.26 mmol), 4-fluoroaniline (0.042 mL, 0.44 mmol) andtriethylamine (0.108 mL, 0.77 mmol) in DCM (2 mL) under nitrogen at 10°C. The resulting solution was stirred at 23° C. for 2 days. A saturatedsolution of sodium carbonate was added and extracted with DCM. Theorganic phase was washed with water, brine, dried over magnesiumsulfate, and concentrated to afford the crude imine. The imine wasdiluted with DCM and MeOH, and acetic acid (0.030 mL, 0.52 mmol) andsodium cyanotrihydroborate (32.4 mg, 0.52 mmol) were added. Theresulting solution was stirred at room temperature for 30 min. Asolution of sodium carbonate was added and extracted with DCM. Theorganic phase was washed with water, brine, dried over magnesiumsulfate, and concentrated to dryness. The crude product was purified byflash chromatography on silica gel eluting with 5% methanol in DCMfollowed by 5% methanolic ammonia (7 N) in DCM. The solvent wasevaporated to dryness and a second purification was done on apreparative HPLC using a Waters X-Terra reverse-phase column (C-18, 5microns silica, 19 mm diameter, 100 mm length, flow rate of 40mL/minute) and decreasingly polar mixtures of water (containing 0.2%ammonium carbonate) and acetonitrile as eluent. The fractions containingthe desired compound were evaporated to dryness to give a white solidwhich was triturated in MTBE then dried under vacuum to affordN-(2-(dimethylamino)ethyl)-8-(1-(4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(73.0 mg, 58.6%). Mass Spectrum: M+H⁺ 483. NMR Spectrum: (CDCl₃) 1.60(d, 3H), 2.25 (s, 6H), 2.50 (t, 2H), 3.45-3.58 (m, 6H), 3.79-3.89 (m,4H), 4.02 (bs, 1H), 4.88-4.98 (m, 1H), 5.56 (s, 1H), 6.40 (dd, 2H), 6.81(dd, 2H), 6.91 (bs, 1H), 8.27 (d, 1H), 8.34 (d, 1H).

The8-acetyl-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamideused as starting material was made as follows:—

Step 1

To a stirred suspension of methyl 4-hydroxybenzoate (180 g, 1183 mmol)in DCM (3 L) was added dropwise bromine (64 mL, 1242 mmol) undernitrogen and at 0° C. and the reaction mixture was left to stir at roomtemperature for 36 hrs. A solution of sodium thiosulfate (500 mL of a10% solution) was then added while keeping the temperature around 15° C.followed by addition of MeOH (250 mL). The organic layer was washed withwater, then brine, dried over magnesium sulfate, filtered andconcentrated to dryness to afford methyl 3-bromo-4-hydroxybenzoate (290g) as a white solid. Mass Spectrum: [M−H]⁻ 229.

Step 2

To a stirred suspension of methyl 3-bromo-4-hydroxybenzoate (270 g, 1168mmol) in DCM (1.5 L) was added pyridine (150 mL). Acetyl chloride (87mL, 1227 mmol) was then added dropwise at room temperature and undernitrogen. The mixture was left to stir for 2 hrs at room temperature.Water (1 L) was then added followed by HCl 2N until pH 1. The organiclayer was then washed with water, brine, dried over magnesium sulfate,filtered and evaporated to dryness to afford methyl4-acetoxy-3-bromobenzoate (300 g, 94%) as a white powder. NMR Spectrum:(DMSOd₆) 2.34 (s, 3H), 3.87 (s, 3H), 7.47 (d, 1H), 8.01 (dd, 1H), 8.20(d, 1H).

Step 3

To methyl 4-acetoxy-3-bromobenzoate (150 g, 549.3 mmol) was addedaluminum trichloride (220 g, 1647.9 mmol) and the mixture was heated at140° C. in the absence of solvent for 3 h. Upon cooling to roomtemperature the solid was crushed and cautiously added to water (1.5 L)with stirring. HCl (250 mL of 12N) was then added and stirring wasmaintained for 30 mins. The solid obtained was collected by filtration,washed with water (2×2 L) and dried overnight to afford3-acetyl-5-bromo-4-hydroxybenzoic acid (120 g, 84%) as a yellow powder.Mass Spectrum: [M−H]⁻ 258.

Step 4

To a stirred suspension of 3-acetyl-5-bromo-4-hydroxybenzoic acid (240g, 926 mmol) in MeOH (2 L) was added dropwise sulfurous dichloride (68mL, 926.5 mmol) under nitrogen and the mixture was heated at 80° C. for3 h. The reaction mixture was cooled to room temperature concentrated,diluted with DCM. The organic layer was washed with brine, dried overmagnesium sulfate, filtered and concentrated to afford a crude compound,which was purified on silica, eluting with 70% of DCM in petroleumether. The solvents were evaporated to dryness to afford methyl3-acetyl-5-bromo-4-hydroxybenzoate (108 g, 42.7%) as a white powder.Mass Spectrum: [M−H]⁻ 229.

Step 5

To a stirred solution of morpholine (201 mL, 2295 mmol) in water (2 L)was added carbon disulfide (0.138 , 2295.67 mmol) under nitrogen. Sodiumhydroxide (96 g, 2410 mmol in solution in 1 of water) was then addeddropwise. The resulting mixture was stirred at room temperature for 1 h,then cooled to 50° C. with an ice bath and dimethyl sulfate (217 mL,2295 mmol) was added dropwise. The mixture was stirred 1 hr at roomtemperature, the obtained solid was collected by filtration, washed withwater (2×1 L) and dried under vacuum over phosphorus pentoxide at 50° C.to give methyl morpholine-4-carbodithioate (360 g, 88%). NMR Spectrum:(CDCl₃): 2.68 (s, 3H), 3.71-3.84 (m, 4H), 4.02 (bs, 2H), 4.30 (bs, 2H).

Step 6

Chlorine gas (455 g, 6417 mmol) was bubbled through a solution of methylmorpholine-4-carbodithioate (170 g, 959 mmol) in DCM (1.5 L) over a 2hrs period, while keeping the temperature around 10-15° C. Once thechlorine addition was completed, stirring was maintained for anadditional 1.5 hr while a precipitation occurred. Nitrogen was thenpassed through the mixture for 30 min. The solid was collected byfiltration under nitrogen, washed with DCM and stored under nitrogen inthe fridge. There was thus obtained 4-(dichloromethylene)morpholin-4-iumchloride (180 g, 92%) as a white hygroscopic solid.

Step 7

To a stirred solution methyl 3-acetyl-5-bromo-4-hydroxybenzoate (106 g,388 mmol) in toluene (1 L) was added dropwise(diethyloxonio)trifluoroborate (0.201 , 1630 mmol), under nitrogen. Theresulting solution was left to stir overnight at room temperature, then4-(dichloromethylene)morpholin-4-ium chloride (143 g, 698 mmol) wasadded and mixture heated at 90° C. for 12 h. Upon cooling to roomtemperature, diethyl ether (1.5 L) was added and the solid was collectedby filtration. This solid was then suspended in MeOH (1 L) and themixture was heated at 50° C. for 2 h.

Upon cooling to room temperature, the solid was collected by filtrationthen solubilized in DCM (1 L) and washed with water and a saturatedsolution of sodium bicarbonate. The organic layer was dried overmagnesium sulfate, filtered and evaporated to dryness to afford methyl8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylate (68.0 g, 47.6%) asan off-white solid. Mass Spectrum: M+H⁺ 368. This intermediate can alsobe prepared by an alternative route (see below)

Step 8

Sodium hydroxide (4.35 mL, 8.69 mmol) was added to a stirred suspensionof methyl 8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylate (1.6 g,4.35 mmol) dissolved in MeOH (30 mL). The resulting suspension wasstirred at 23° C. for 16 hrs. The mixture was diluted with water and thepH adjusted to 3 with HCl 2N. The precipitate was collected byfiltration, washed with water and dried over phosphorus pentoxideovernight to give 8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (1.30 g, 84%) as a beige solid. Mass Spectrum: M+H⁺ 356

Step 9

Bis(triphenylphosphine) palladium (II) chloride (12.78 mg, 0.02 mmol)was added to a stirred mixture of8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (215 mg, 0.61mmol) and tributyl(1-ethoxyvinyl)stannane (0.226 mL, 0.67 mmol) in1,4-dioxane (5 mL) and the mixture was purged with nitrogen. Theresulting mixture was stirred at 100° C. for 3 hrs. HCl 2N (0.5 mL) wasadded and the reaction mixture was stirred at 50° C. for 25 minutes thenallowed to cool to room temperature and concentrated under vacuum. Theproduct was diluted with diethyl ether and pentane to afford a solid,which was collected by filtration and washed with ether/pentane to give8-acetyl-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (200 mg, 104%)as a beige solid. Mass Spectrum: M+H⁺318.

Step 10

To a suspension of 8-acetyl-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (470 mg, 1.48 mmol) in DCM (10 mL) at room temperature, were addedto DIPEA (0.284 mL, 1.63 mmol) and TSTU (491 mg, 1.63 mmol). The mixturewas stirred for 15 minutes, N1,N1-dimethylethane-1,2-diamine (0.171 mL,1.56 mmol) was added stirring was maintained for an additional hour. Themixture was adsorbed on silica gel and purified by flash chromatographyon silica gel eluting with 5 to 10% methanol in DCM then with 10%methanolic ammonia (7 N) in DCM. The solvent was evaporated to drynessto afford8-acetyl-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(352 mg, 61.3%) as a beige solid. Mass Spectrum: M+H⁺ 388.

Alternative route to prepare methyl8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylate:

Step 1

Dibromine (0.185 L, 3614.92 mmol) was added dropwise a stirredsuspension of methyl 4-hydroxybenzoate (500 g, 3286 mmol) in DCM (4 L)at 0° C. under N2. The mixture was left to stir for 24 hrs at RT underN2 (need to trap HBr). A solution of sodium metabisulfite (62.5 g, 329mmol) in 2 L of water was then added, while keeping the temperaturearound 15° C., followed by 500 mL of MeOH. The organic layer was washedwith water, brine, dried over magnesium sulfate, filtered andconcentrated to dryness to afford methyl 3-bromo-4-hydroxybenzoate (710g, 94%) as a white solid. NMR Spectrum (CDCl3): 3.89 (s, 3H), 5.95 (s,1H), 7.05 (d, 1H), 7.92 (dd, 1H), 8.19 (d, 1H).

Step 2

To a degassed solution of methyl 3-bromo-4-hydroxybenzoate (350 g,1514.87 mmol) in ethanol (3 L) were added triethylamine (0.528 L,3787.17 mmol), 1-(vinyloxy)butane (0.588 L, 4544.60 mmol), 1,1′-bis(diphenylphosphino)ferrocene (33.1 g, 60.6 mmol) and diacetoxypalladium(8.50 g, 37.9 mmol) under nitrogen. The mixture was heated at 70° C.overnight. The reaction was cooled down, filtered and the filtrateconcentrated. The resulting solid was solubilized with DCM (2 L) and HCl4N (1.14 L, 4544 mmol) was added under stirring. Stirring was maintainedfor 2 hrs, the organic phase was separated, dried over magnesiumsulfate, filtered and concentrated to afford a solid which was stirredin diethyl ether (5 L) for 2 hrs. The solid was filtered off and thefiltrate concentrated to dryness to afford methyl3-acetyl-4-hydroxybenzoate (240 g, 82%) as a beige powder. Massspectrum: [M−H]⁻ 193.

Step 3

To a stirred solution of methyl 3-acetyl-4-hydroxybenzoate (240 g, 1236mmol) in DCM (2 L) was added pyridine (0.400 L, 4944 mmol) followed by adropwise addition of dibromine (0.070 L, 1360 mmol) at 0° C. Thereaction mixture was stirred at RT for 2 hrs then cooled to 5° C. andHCl 4N (0.927 L, 3708 mmol) was added dropwise. The organic phase wasseparated, dried over magnesium sulfate, filtered and concentrated toafford a brown solid which was stirred in ether/petroleum ether (1:1, 1L) for 1 hr. The solid was collected by filtration and dried to affordmethyl 3-acetyl-5-bromo-4-hydroxybenzoate (270 g, 80%) as a beigepowder.

Step 4

To a solution of lithium bis(trimethylsilyl)amide (1.41 L, 1406 mmol) at−65° C. under nitrogen was added dropwise methyl3-acetyl-5-bromo-4-hydroxybenzoate (120 g, 439 mmol) in THF (1.2 L). Thesolution was allowed to warm to 0° C., and maintained at thistemperature for 1 hr. The solution was cooled back to −65° C. andmorpholine-4-carbonyl chloride (0.055 L, 483 mmol) was added. Themixture was stirred at RT for 2 hrs then cooled to −30° C., DCM (1.5 L)and water (1 L) were added followed by dropwise addition of HCl 6N (500mL) then HCl 2N (300 mL) until pH 7, the aqueous solution was extractedwith DCM (3×). The combined extracts were dried over magnesium sulfateand evaporated. The crude product was triturated in MTBE to obtainmethyl 3-bromo-4-hydroxy-5-(3-morpholino-3-oxopropanoyl)benzoate (153 g,90%) as a beige solid. Mass Spectrum: M+H⁺ 388.

Step 5

Trifluoromethanesulfonic anhydride (0.755 L, 4487 mmol) was added to astirred solution of methyl3-bromo-4-hydroxy-5-(3-morpholino-3-oxopropanoyl)benzoate (433 g, 1122mmol, pooled material from several batches) dissolved in1,2-dichloroethane (1 L) at RT under nitrogen (exotherm). The resultingsolution was stirred at 50° C. overnight. The mixture was partiallyevaporated, and the residue was diluted with MeOH (1.6 L) at 0° C.(exotherm) and stirred for 1 hr at RT. The solvent was evaporated againand the residue was diluted in DCM, quenched with a saturated aqueoussolution of sodium carbonate and extracted with DCM. The combinedorganic phases were washed with brine, dried over magnesium sulfate andconcentrated to afford the crude product. The crude was triturated underMTBE (2×), ethyl acetate (1×) and MTBE (1×). The solid was dried toafford methyl 8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylate (208g, 50%) as a beige solid. Mass Spectrum: M+H⁺ 370.

EXAMPLE 1.01N-(2-(dimethylamino)ethyl)-8-(1-(3-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

N-(2-(dimethylamino)ethyl)-8-(1-(3-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamidewas prepared using an analogous procedure to that described in Example1.00 (28.0 mg, 18.7%). Mass Spectrum: M+H⁺ 483. NMR Spectrum: (CDCl₃)1.62 (d, 3H), 2.27 (s, 6H), 2.52 (t, 2H), 3.44-3.58 (m, 6H), 3.78-3.89(m, 4H), 4.55 (bs, 1H), 4.92-5.02 (m, 1H), 5.56 (s, 1H), 6.14 (ddd, 1H),7.27 (dd, 1H), 7.36 (ddd, 1H), 6.95 (bs, 1H), 7.03 (dd, 1H), 8.28 (d,1H), 8.35 (d, 1H).

EXAMPLE 1.02N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-8-(1-(phenylamino)ethyl)-4H-chromene-6-carboxamide

N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-8-(1-(phenylamino)ethyl)-4H-chromene-6-carboxamidewas prepared using an analogous procedure to that described in Example1.00 (36.0 mg, 25%). Mass Spectrum: M+H⁺ 465. NMR Spectrum: (CDCl₃) 1.62(d, 3H), 2.25 (s, 6H), 2.49 (t, 2H), 3.44-3.57 (m, 6H), 3.75-3.86 (m,4H), 4.09 (d, 1H), 4.95-5.05 (m, 1H), 5.56 (s, 1H), 6.58 (d, 2H), 6.68(t, 1H), 6.90 (bs, 1H), 7.11 (dd, 2H), 8.30 (d, 1H), 8.34 (d, 1H).

EXAMPLE 2.008-(1-(3-chloro-2-fluorophenylamino)ethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (73.3 mg, 0.23 mmol) was added to a stirred solutionof8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (85 mg, 0.19 mmol), 4-methylmorpholine (0.052 mL, 0.48 mmol) andN1,N1-dimethylethane-1,2-diamine (0.025 mL, 0.23 mmol) dissolved in NMP(1.2 mL) at room temperature. The resulting solution was stirred for 2hrs. The reaction mixture was purified by preparative HPLC using aWaters SunFire reverse-phase column (C-18, 5 microns silica, 19 mmdiameter, 100 mm length, flow rate of 40 mL/minute) and decreasinglypolar mixtures of water (containing 0.2% ammonium carbonate) andacetonitrile as eluent. The fractions containing the desired compoundwere evaporated, triturated with diethyl ether and dried to afford8-(1-(3-chloro-2-fluorophenylamino)ethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(59.2 mg, 60.2%) as a white solid. Mass Spectrum: M+H⁺ 517. NMRSpectrum: (DMSOd₆) 1.57 (d, 3H), 2.15 (s, 6H), 2.37 (t, 2H), 3.26-3.34(m partially hidden by H2O, 2H), 3.50-3.62 (m, 4H), 3.70-3.79 (m, 4H),5.03-5.12 (m, 1H), 5.61 (s, 1H), 6.34 (dd, 1H), 6.49 (d, 1H), 6.64 (ddd,1H), 6.82 (d, 1H), 8.10 (d, 1H), 8.32 (d, 1H), 8.61 (t, 1H).

The8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid used as starting material was made as follows:—

To a stirred suspension of methyl8-bromo-2-morpholino-4-oxo-4H-chromene-6-carboxylate (40 g, 108 mmol, asdescribed in Example 1.00) in dioxane (300 mL) was addedtributyl(1-ethoxyvinyl)stannane (38.5 mL, 114 mmol) andbis(triphenylphosphine) palladium (II) chloride (3.05 g, 4.35 mmol). Themixture was purged with nitrogen and heated at 90° C. overnight. Moretributyl(1-ethoxyvinyl)stannane (20 mL) and bis(triphenylphosphine)palladium (II) chloride (1.5 g) were added and the reaction mixtureheated for an additional 3 hrs. The mixture was cooled to roomtemperature, HCl 2N (81 mL, 163 mmol) was added and the dark suspensionheated at 45° C. for 30 min. The dioxane was evaporated under vacuum,the residue was dissolved in DCM and a saturated solution of sodiumbicarbonate was added until pH 4. The organic layer was washed withbrine, dried over magnesium sulfate, filtered and concentrated to afforda crude compound which was triturated in diethylether, filtered anddried to afford methyl8-acetyl-2-morpholino-4-oxo-4H-chromene-6-carboxylate (25.0 g, 69.5%) asan off-white solid. Mass Spectrum: M+H⁺ 332.

Sodium tetrahydroborate (82 mg, 2.16 mmol) was added to a solution ofmethyl 8-acetyl-2-morpholino-4-oxo-4H-chromene-6-carboxylate (650 mg,1.96 mmol) in methanol (20 mL) and DCM (10 mL) at −10° C. After 15 minof stirring at −10° C., the reaction mixture was quenched with water (25mL). The volatiles were removed and the aqueous layer extracted twicewith DCM. The combined organics phases were washed with brine, driedover magnesium sulfate and concentrated. The residue was triturated withdiethylether and the solid collected by filtration to give methyl8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate (500 mg,76%) as a brown solid, which used for the next step without furtherpurification. Mass Spectrum: M+H⁺ 334.

Tribromophosphine (1.65 mL, 1.65 mmol) was added to a stirred suspensionof methyl8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate (500 mg,1.50 mmol) in DCM (10 mL) in an ice bath under nitrogen. The resultingsolution was stirred at room temperature for 24 hrs. The reaction wasincomplete and further tribromophosphine (0.300 mL, 0.30 mmol) was addedand the reaction mixture stirred for an additional 12 hrs. The solventwas evaporated, the residue was suspended in water and ice and a sodiumcarbonate solution was carefully added until pH 6. The precipitate wascollected by filtration, washed with water then with diethylether anddried to afford methyl8-(1-bromoethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate (470 mg,79%) as a dark beige solid. Mass Spectrum: M+H⁺ 398.

3-chloro-2-fluoroaniline (0.239 mL, 2.17 mmol) was added to a stirredsuspension of methyl8-(1-bromoethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate (215 mg,0.54 mmol) dissolved in DCM (3 mL) at room temperature. The resultingsuspension was stirred for 16 hrs then the temperature was increased to50° C. for 16 hrs. The crude product was purified by flashchromatography on silica gel eluting with 5% methanol in DCM. Thesolvent was evaporated to dryness to afford methyl8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate(206 mg, 82%) as a white solid. Mass Spectrum: M+H⁺ 461.

Sodium hydroxide (0.217 mL, 0.43 mmol) was added to a stirred solutionof methyl8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate(200 mg, 0.43 mmol) dissolved in MeOH (2 mL). The resulting solution wasstirred at room temperature for 16 hrs. THF was added and the resultingsolution was stirred at 50° C. for 8 hrs. The pH was adjusted to 3 withHCl 2N, the solvent was removed under vacuum and the residue dilutedwith H2O. The solid was collected by filtration, washed with water anddried to give 8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (171 mg, 88%) asa white solid. Mass Spectrum: M+H⁺ 447.

EXAMPLE 2.018-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

8-(1-(4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (82 mg, 0.20mmol) was reacted with dimethylamine (0.119 mL, 0.24 mmol) using aprocedure similar to the one described in Example 2.00 to afford8-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(53 mg, 60.7%) as a white solid. Mass Spectrum: M+H⁺ 440.NMR Spectrum:(DMSOd₆) 1.50 (d, 3H), 2.66 (s, 3H), 2.92 (s, 3H), 3.50-3.65 (m, 4H),3.70-3.81 (m, 4H), 4.92-5.02 (m, 1H), 5.60 (s, 1H), 6.27 (d, 1H), 6.46(dd, 2H), 6.84 (dd, 2H), 7.56 (d, 1H), 7.77 (d, 1H).

The 8-(1-(4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid used as startingmaterial was made using a procedure similar to the one described inExample 2.00. Mass Spectrum: M+H⁺ 413.

EXAMPLE 2.02N-(2-(dimethylamino)ethyl)-8-(1-(4-fluorophenylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

8-(1-(4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (80 mg, 0.19mmol) was reacted with N1,N1,N2-trimethylethane-1,2-diamine (0.030 mL,0.23 mmol) using a procedure similar to the one described in Example2.00 to affordN-(2-(dimethylamino)ethyl)-8-(1-(4-fluorophenylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(15.0 mg, 15.6%) as a white solid. Mass Spectrum: M+H⁺497. NMR Spectrum:(DMSOd₆) 1.49 (d, 3H), 1.74 (bs, 3H), 2.01 (bs, 1H), 2.18 (s, 3H), 2.66(bs, 1.5H), 2.89 (bs, 2H), 3.05 (bs, 1.5H), 3.47 (bs, 1H), 3.50-3.64 (m,4H), 3.71-3.79 (m, 4H), 4.93-5.01 (m, 1H), 5.59 (s, 1H), 6.29 (bs, 1H),6.46 (dd, 2H), 6.84 (dd, 2H), 6.54 (d, 1H), 6.73 (d, 1H).

EXAMPLE 2.038-(1-(3-chloro-4-fluorophenylamino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one

8-(1-(3-chloro-4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (125 mg, 0.28 mmol) was reacted with piperidin-4-ol (34.0 mg, 0.34mmol) using a procedure similar to the one described in Example 2.00 toafford8-(1-(3-chloro-4-fluorophenylamino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(110 mg, 74.2%) as a white solid. Mass Spectrum: M+H⁺ 530. NMR Spectrum:(DMSOd₆) at 323° K: 1.24 (bs, 2H), 1.51 (d, 3H), 2.99 (bs, 2H), 3.30(bs, 2H), 3.37 (bs, 2H), 3.49-3.63 (m, 4H), 3.63-3.72 (m, 1H), 3.72-3.82(m, 4H), 4.62 (d, 1H), 4.92-5.02 (m, 1H), 5.56 (s, 1H), 6.36-6.46 (m,2H), 6.59 (dd, 1H), 7.01 (dd, 1H), 7.51 (d, 1H), 7.76 (d, 1H).

The8-(1-(3-chloro-4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid used as starting material was made using a procedure similar to theone described in Example 2.00. Mass Spectrum: M+H⁺ 447.

EXAMPLE 2.048-(1-(3-chloro-4-fluorophenylamino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

8-(1-(3-chloro-4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (78 mg, 0.17 mmol) was reacted with 2-(methylamino)ethanol (0.017mL, 0.21 mmol) using a procedure similar to the one described in Example2.00 to give8-(1-(3-chloro-4-fluorophenylamino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(60.0 mg, 68.2%) as a white solid. Mass Spectrum: M+H⁺504. NMR Spectrum:(DMSOd₆) at 323° K: 1.51 (d, 3H), 2.82 (bs, 1.5H), 2.90 (bs, 1.5H), 3.42(bs, 4H), 3.49-3.63 (m, 4H), 3.69-3.79 (m, 4H), 4.60 (bs, 1H), 4.92-5.01(m, 1H), 5.56 (s, 1H), 6.37 (dd, 1H), 6.42 (ddd, 1H), 6.61 (dd, 1H),7.02 (dd, 1H), 7.57 (d, 1H), 7.80 (d, 1H).

The8-(1-(3-chloro-4-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid used as starting material was made using a procedure similar to theone described in Example 2.0. Mass Spectrum: M+H⁺ 447.

EXAMPLE 2.058-(1-(3-chloro-2-fluorophenylamino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (130 mg, 0.29 mmol) was reacted with 2-(methylamino)ethanol (0.028mL, 0.35 mmol) using a procedure similar to the one described in Example2.00 to afford8-(1-(3-chloro-2-fluorophenylamino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(65.0 mg, 44.3%) as a white solid. Mass Spectrum: M+H⁺504. NMR Spectrum:(DMSOd₆) 1.57 (d, 3H), 2.76 (s, 1.5H), 2.94 (s, 1.5H), 3.07 (bs, 1H),3.27 (bs, 1H), 3.45 (bs, 1H), 3.51-3.64 (m, 5H), 3.70-3.78 (m, 4H), 4.69(bs, 0.5H), 4.78 (ns, 0.5H), 5.08 (bs, 1H), 5.60 (s, 1H), 6.33-6.47 (bs,2H), 6.65 (ddd, 1H), 6.83 (dd, 1H), 7.60 (bs, 0.5H), 7.61 (bs, 0.5H),7.79 (bs, 0.5H), 7.81 (bs, 0.5H).

The8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid used as starting material was made using a procedure similar to theone described in Example 2.00. Mass Spectrum: M+H⁺ 447.

EXAMPLE 2.068-(1-(3-chloro-2-fluorophenylamino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one

8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (130 mg, 0.29 mmol) was reacted with piperidin-4-ol (35.3 mg, 0.35mmol) using a procedure similar to the one described in Example 2.00 togive8-(1-(3-chloro-2-fluorophenylamino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(101 mg, 65.5%) as a white solid. Mass Spectrum: M+H⁺ 530. NMR Spectrum:(DMSOd₆) at 323° K: 1.24 (bs, 2H), 1.62 (d, 3H), 1.63 (bs, 2H), 2.99(bs, 2H), 3.19 (bs partially hidden by H2O, 2H), 3.49-3.63 (m, 4H),3.65-3.72 (m, 1H), 3.72-3.78 (m, 4H), 4.62 (d, 1H), 5.04-5.13 (m, 1H),5.56 (s, 1H), 6.28 (d, 1H), 6.33 (dd, 1H), 6.64 (ddd, 1H), 6.82 (ddd,1H), 7.57 (d, 1H), 7.77 (d, 1H).

EXAMPLE 2.07 8-(1-(3,5-difluorophenylamino)ethyl)-6-(morpholine-4-carbonyl)-2-morpholino-4H-chromen-4-one

TSTU (84 mg, 0.28 mmol) at 25° C., was added to8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (100 mg, 0.14 mmol) and DIPEA (0.049 mL, 0.28 mmol) dissolved inDCM (1 mL). The resulting solution was stirred at 25° C. for 2 hrs.Morpholine (0.037 mL, 0.42 mmol) was then added, the resulting solutionwas stirred at 25° C. for 30 minutes then concentrated. The crude wasdiluted with 1 mL of DMA and purified by preparative HPLC using a WatersX-Bridge reverse-phase column (C-18, 5 microns silica, 19 mm diameter,100 mm length, flow rate of 40 mL/minute) and decreasingly polarmixtures of water (containing 0.2% ammonium carbonate) and acetonitrileas eluent. The fractions containing the desired compound were evaporatedto dryness, triturated in diethyl ether and collected by filtration toafford 8-(1-(3,5-difluorophenylamino)ethyl)-6-(morpholine-4-carbonyl)-2-morpholino-4H-chromen-4-one (63.0 mg,90%) as a yellow solid. Mass Spectrum: M+H⁺ 500. NMR Spectrum: (DMSOd₆)1.53 (d, 3H), 3.11 (bs, 2H), 3.37-3.71 (m, 10H), 3.71-3.80 (m, 4H),4.98-5.07 (m, 1H), 5.62 (s, 1H), 6.15 (dd, 2H), 6.25 (ddd, 1H), 6.97 (d,1H), 7.51 (d, 1H), 7.82 (d, 1H).

The8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid used as starting material was made using a procedure similar to theone described in Example 2.00.

EXAMPLE 2.088-(1-(3,5-difluorophenylamino)ethyl)-6-((S)-3-hydroxypyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(enantiomer 2)

EDCI (102 mg, 0.53 mmol) was added in one portion to8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (200 mg, 0.35mmol, enantiomer 2 ([α]^(D) _(20°) −102°, described as a startingmaterial in Example 7.0a), (S)-pyrrolidin-3-ol (0.043 mL, 0.53 mmol) andHOPO (47.1 mg, 0.42 mmol) dissolved in DCM (2 mL) in a screwcap vial.The resulting solution was stirred at RT for 5 min then 50° C. for 30min. More EDCI (˜50 mg) was added to complete the reaction. The solutionwas washed with a 10% aq. citric acid solution, water, brine and driedover magnesium sulfate. The solvent was evaporated and the crude productpurified by flash chromatography on silica gel (40 g) eluting with 0 to10% MeOH in DCM. The solvent was evaporated to dryness to afford8-(1-(3,5-difluorophenylamino)ethyl)-6-((S)-3-hydroxypyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(95%) as a off-white foam. Mass Spectrum: M+H⁺ 500. NMR Spectrum(CDCl3): 1.60 (d, 3H), 1.89-2.11 (m, 1H), 2.54 (bs, 0.5H), 2.81 (bs,0.5H), 3.29-3.43 (m, 2H), 3.51 (bs, 4H), 3.64-3.78 (m, 2H), 3.79-3.91(m, 5H), 4.42 (bs, 0.5H), 4.54 (bs, 0.5H), 4.59 (bs, 0.5H), 4.70 (bs,0.5H), 4.87-4.95 (m, 1H), 5.54 (bs, 1H), 5.89-6.01 (m, 2H), 6.09 (dd,1H), 7.85 (s, 0.5H), 7.87 (s, 0.5H), 8.18 (bs, 0.5H), 8.25 (bs, 0.5H).

EXAMPLE 3.00N-(2-(dimethylamino)ethyl)-8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

A solution of8-(1-bromoethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (85 mg, 0.16 mmol) and 4-fluoro-N-methylaniline (0.077 mL,0.64 mmol) in NMP (1 mL) was stirred at room temperature for 4 h then at45° C. for 1 h. The reaction mixture was allowed to cool to roomtemperature and purified by preparative HPLC using a Waters X-Bridgereverse-phase column (C-18, 5 microns silica, 19 mm diameter, 100 mmlength, flow rate of 40 mL/minute) and decreasingly polar mixtures ofwater (containing 0.2% ammonium carbonate) and acetonitrile as eluent.The fractions containing the desired compound were evaporated todryness. The residue was triturated in diethyl ether, collected byfiltration and dried to affordN-(2-(dimethylamino)ethyl)-8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(42.0 mg, 53.1%) as a white solid. Mass Spectrum: M+H⁺ 497. NMRSpectrum: (CDCl₃) 1.66 (d, 3H), 2.33 (s, 6H), 2.33 (bs, 2H), 2.60 (bs,2H), 2.61 (s, 3H), 3.09-3.24 (m, 4H), 3.44-3.57 (m, 4H), 3.57-3.66 (m,2H), 5.36 (q, 1H), 5.48 (s, 1H), 6.75 (dd, 2H), 6.77 (dd, 2H), 7.14 (bs,1H), 8.33 (s, 1H), 8.44 (s, 1H).

The8-(1-bromoethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide used as starting material was made as follows:—

To a suspension of methyl8-acetyl-2-morpholino-4-oxo-4H-chromene-6-carboxylate (193 mg, 0.41mmol, as described in Example 2.00) in methanol (2 mL) was added at −15°C. sodium tetrahydroborate (15.4 mg, 0.41 mmol). The resultingsuspension was stirred at −15° C. for 20 minutes. The reaction mixturewas quenched with a 2N aqueous NaOH solution (0.408 mL, 0.82 mmol) andallowed to warm to room temperature for 1 hr. More NaOH (0.408 mL, 0.82mmol) was added and stirring was maintained for another 15 min. HCl(0.917 mL, 1.83 mmol) was added to adjust pH to 2-3. The resultingprecipitate was diluted with a about 1 mL of water, collected byfiltration, washed with ethyl acetate, diethyl ether and dried to aconstant weight to afford8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (122mg, 94%) as a orange solid which was used without further purification.Mass Spectrum: M+H⁺ 320.

TSTU (104 mg, 0.34 mmol) at 25° C., was added portionwise to8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (110mg, 0.34 mmol) and DIPEA (0.066 mL, 0.38 mmol) suspended in DCM (1 mL)under nitrogen. The resulting mixture was stirred at 25° C. for 1.5 hrs.N1,N1-dimethylethane-1,2-diamine (0.038 mL, 0.34 mmol) was then addedand the mixture stirred at 25° C. for 30 minutes. The mixture was pouredonto a silica gel column and purified by flash chromatography elutingwith 5% methanolic ammonia (7 N) in DCM. The solvent was evaporated todryness, the residue was triturated in diethyl ether, collected byfiltration and dried to affordN-(2-(dimethylamino)ethyl)-8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(80 mg, 0.205 mmol, 59.6%) as a off-white solid. Mass Spectrum: M+H⁺390.

A solution of tribromophosphine 1M in DCM (0.154 mL, 0.15 mmol) at 25°C., was added dropwise toN-(2-(dimethylamino)ethyl)-8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(60 mg, 0.15 mmol) suspended in DCM (1 mL). This suspension was stirredat 25° C. for 4 days. The resulting precipitate was collected byfiltration, washed with diethyl ether and dried to afford8-(1-bromoethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (98 mg, >100%) as a white solid, which was used withoutfurther purification. Mass Spectrum: M+H⁺ 454.

EXAMPLE 3.018-(1-((3,4-difluorophenyl)(methyl)amino)ethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3,4-difluoro-N-methylaniline (161 mg, 1.13 mmol) was reacted with8-(1-bromoethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (150 mg, 0.28 mmol) using a procedure similar to the onedescribed in Example 3.00 to give8-(1-((3,4-difluorophenyl)(methyl)amino)ethyl)-N-(2-(dimethylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(66.0 mg, 45.6%). Mass Spectrum: M+H⁺ 515. NMR Spectrum: (DMSOd₆) 1.56(d, 3H), 2.18 (s, 6H), 2.41 (t, 2H), 2.63 (s, 3H), 3.21-3.28 (m, 2H),3.34-3.41 (m partially hidden by H2O, 4H), 3.41-3.49 (m, 2H), 3.49-3.56(m, 2H), 5.53 (q, 1H), 5.55 (s, 1H), 6.61 (d, 1H), 6.87 (ddd, 1H), 7.24(dd, 1H), 8.09 (d, 1H), 8.43 (d, 1H), 8.74 (t, 1H).

EXAMPLE 3.02N-(2-(dimethylamino)ethyl)-8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

4-fluoro-N-methylaniline (82 mg, 0.66 mmol) was reacted with8-(1-bromoethyl)-N-(2-(dimethylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (90 mg, 0.16 mmol) using a procedure similar to the onedescribed in example 3.00 to giveN-(2-(dimethylamino)ethyl)-8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(39.0 mg, 46.4%) as a white solid. Mass Spectrum: M+H⁺ 511. NMRSpectrum: (DMSOd₆) at 323° K: 1.55 (d, 3H), 2.04 (bs, 6H), 2.40 (bs,2H), 2.68 (s, 3H), 2.93 (s, 3H), 3.22-3.36 (m, 4H), 3.37 (bs, 2H),3.44-3.58 (m, 4H), 5.45 (q, 1H), 5.50 (s, 1H), 6.82 (dd, 2H), 7.01 (dd,2H), 7.56 (d, 1H), 7.83 (d, 1H).

The8-(1-bromoethyl)-N-(2-(dimethylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide used as starting material was made using a proceduresimilar to the one described for the starting material in Example 3.00except that N1,N1,N2-trimethylethane-1,2-diamine was used instead ofN1,N1-dimethylethane-1,2-diamine.

EXAMPLE 3.038-(1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3,4-difluoroaniline (0.105 mL, 1.06 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (130 mg, 0.27 mmol) using a procedure similar to the onedescribed in Example 3.00 to give8-(1-(3,4-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(35.0 mg, 28.8%) as a white solid. Mass Spectrum: M+H⁺ 458. NMRSpectrum: (DMSOd₆) 1.50 (d, 3H), 2.70 (s, 3H), 2.94 (s, 3H), 3.50-3.63(m, 4H), 3.71-3.79 (m, 4H), 4.92-5.01 (m, 1H), 5.60 (s, 1H), 6.25 (d,1H), 6.46 (ddd, 1H), 6.54 (d, 1H), 7.05 (dd, 1H), 7.54 (d, 1H), 7.78 (d,1H).

A larger batch of the above racemic compound was resolved by chiralpreparative HPLC using the following conditions:

Column CelluCoat 250 × 50 10 μm Eluent Heptane/EtOH/TEA 50/50/0.1 OvenTemperature Ambient Flow 118 mL/min Wavelength 300 nm Sample Conc 50mg/ml Heptane/EtOH 1/1 Injection amount 600 mg

2.19 g of racemic compound was separated using the above conditions togive:

First eluting enantiomer 1100 mg (Example 3.03a) [α]^(D) _(20°): +119°(99.1% ee) in MeCN

Second eluting enantiomer 1090 mg (Example 3.03b) [α]^(D) _(20°): −120°(99.3% ee) in MeCN

The8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide used as starting material was made as follows:

TSTU (108 mg, 0.36 mmol) was added portionwise to8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (115mg, 0.36 mmol, as described in Example 3.00) and DIPEA (0.069 mL, 0.40mmol) suspended in DCM (1 mL) at 25° C. under nitrogen. The resultingmixture was stirred at 25° C. for 1.5 hrs. Dimethylamine (0.180 ml, 0.36mmol) was then added and stirring was maintained for an additional 30min. The mixture was poured onto a silica gel column and purified byflash chromatography eluting with 5% methanolic ammonia (7 N) indichloromethane. The solvent was evaporated to dryness. The residue wastriturated in diethyl ether, collected by filtration and dried to afford8-(1-hydroxyethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(123 mg, 0.355 mmol, 99%) as a off-white solid. Mass Spectrum: M+H⁺ 347.

A solution of tribromophosphine 1M in CH2Cl2 (0.346 mL, 0.35 mmol) at25° C., was added dropwise to8-(1-hydroxyethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(120 mg, 0.35 mmol) suspended in DCM (1 mL). The resulting suspensionwas stirred at 25° C. for 4 days. The resulting gum was triturated inether to give a precipitate which was collected by filtration, washedwith ether and dried to a constant weight to afford8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (140 mg, 82%) as a white solid, which was used withoutfurther purification. Mass Spectrum: M+H⁺ 409.

EXAMPLE 3.048-(1-(3-chloro-2-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-chloro-2-fluoroaniline (0.179 mL, 1.63 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (200 mg, 0.41 mmol, as described in Example 3.03) using aprocedure similar to the one described in Example 3.00 to give8-(1-(3-chloro-2-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(121 mg, 62.6%) as a white solid. Mass Spectrum: M+H⁺ 474. NMR Spectrum:(DMSOd₆) 1.58 (d, 3H), 2.70 (s, 3H), 2.94 (s, 3H), 3.50-3.64 (m, 4H),3.70-3.80 (m, 4H), 5.05-5.15 (m, 1H), 5.60 (m, 1H), 6.37 (dd, 1H), 6.44(d, 1H), 6.65 (ddd, 1H), 6.83 (dd, 1H), 7.60 (d, 1H), 7.78 (d, 1H).

A larger batch of the above racemic compound was resolved by chiralpreparative HPLC using the following conditions:

Column Chiralpak IC 21 × 250 mm, 5 μm Eluent DCM/isopropanol 1:1 OvenTemperature Ambient Flow 20 mL/min Wavelength 220 nm Sample Conc 50mg/mL in DCM/MeOH 1:1 Injection 100 mg

1.8 g of racemic compound was separated using the above conditions togive: First eluting enantiomer 741 mg (Example 3.04a) [α]^(D) _(20°):+159° (>98% ee) in MeCN Second eluting enantiomer 622 mg (Example 3.04b)[α]^(D) _(20°): −159° (>98% ee) in MeCN

EXAMPLE 3.058-(1-(3-chloro-4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-chloro-4-fluoroaniline (238 mg, 1.63 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (200 mg, 0.41 mmol, as described in Example 3.03) to give8-(1-(3-chloro-4-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(122 mg, 63.1%) as a white solid. Mass Spectrum: M+H⁺ 474. NMR Spectrum:(DMSOd₆) 1.51 (d, 3H), 2.70 (s, 3H), 2.94 (s, 3H), 3.50-3.64 (m, 4H),3.69-3.81 (m, 4H), 4.94-5.05 (m, 1H), 5.60 (m, 1H), 6.47 (ddd, 1H), 6.52(d, 1H), 6.67 (dd, 1H), 7.05 (dd, 1H), 7.55 (d, 1H), 7.79 (d, 1H).

EXAMPLE 3.068-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

To a suspension of8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (3.63 g, 7.41 mmol, as described in Example 3.03) in DMF(35 mL) under nitrogen was added 3,5-difluoroaniline (3.82 g, 29.62mmol). The resulting yellow solution was stirred at 50° C. for 5 hrs.The reaction mixture was concentrated to dryness, then purified by flashchromatography on silica gel eluting with 1 to 7% methanol in DCM. Thesolvent was evaporated to dryness to afford8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(2.32 g, 68.5%) as a pale yellow solid. Mass Spectrum: M+H⁺ 458. NMRSpectrum: (DMSOd₆) 1.52 (d, 3H), 2.74 (s, 3H), 2.95 (s, 3H), 3.50-3.64(m, 4H), 3.70-3.79 (m, 4H), 4.97-5.05 (m, 1H), 5.60 (m, 1H), 6.15 (dd,2H), 6.22 (tt, 1H), 6.96 (d, 1H), 7.54 (d, 1H), 7.81 (d, 1H).

This racemic compound was resolved by chiral preparative HPLC using thefollowing conditions:

Instrument Gilson Prep (200 mL heads) Column Merck 50 mm 20 μm ChiralpakIC Eluent MeCN/MeOH/DEA 90/10/0.2 Oven Temperature Ambient Flow 60mL/min Wavelength 254 nm Sample Conc 12 mg/mL in MeCN/MeOH/DEA 90/10/0.5Injection volume 30 mL Run Time 50 minFirst eluting enantiomer (r.t: 10.8 min) 0.820 g (Example 3.06a) [α]^(D)₂₀=+121.8° in EtOH.Second eluting enantiomer (r.t: 15.4 min) 0.923 g (Example 3.06b)[α]^(D) ₂₀=−122.6° in EtOH.Retention times (r.t.) are from analytical HPLC post chiral separation(1 mL/min, 20 μm Chiralpak AD MeCN/MeOH/DEA 90/10/0.5).

The Second eluting enantiomer (Example 3.06b) was crystallized inethanol, before drying under ambient conditions to yield Form Amaterial. This form was determined to be crystalline by XRPD (seeFIG. 1) and had the following characteristic X-Ray Powder Diffractionpeaks:

Angle 2- Theta (2θ) Intensity % 7.9 100.0 16.7 9.2 20.3 8.0 19.3 7.713.2 7.3 7.2 6.8 19.5 6.4 17.9 5.8 23.0 5.8 5.0 5.5DSC analysis of Form A was also carried out (FIG. 2) and showed aninitial event with an onset at 125.8° C. and a peak at 129.2° C.,followed by an exothermic event before a melt with an onset of 223.8° C.and a peak at 226.7° C.

Form B material was produced by slurrying Form A material inacetonitrile, ethylacetate or methanol. With each of the given solventsapproximately 20 mg of the original material was placed in a vial with amagnetic flea, and approximately 2 ml of solvent added, the vial wasthen sealed tightly with a cap and left to stir on a magnetic stirrerplate. After 3 days, the sample was removed from the plate, the captaken off and the slurry left to dry under ambient conditions before itwas analysed by XRPD and DSC. This form (Form B) was determined to becrystalline by XRPD (FIG. 3) and seen to be different to Form A. Thisform had the following characteristic X-Ray Powder Diffraction peaks:

Angle 2- Theta (2θ) Intensity % 20.7 100 13.8 54.7 21.5 50.8 19.6 36.612.8 35.7 15.4 24.9 10.7 20.5 8.5 19.7 22.4 18.9DSC analysis (FIG. 4) showed that the Form B material had a meltingpoint of 225.8° C. (onset).

EXAMPLE 3.078-(1-((4-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

4-fluoro-N-methylaniline (204 mg, 1.63 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (200 mg, 0.41 mmol, as described in Example 3.03), using ananalogous procedure to that described in Example 3.00, to afford8-(1-((4-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(96 mg, 51.9%) as a white solid. Mass Spectrum: M+H⁺ 454. NMR Spectrum:(DMSOd₆) 1.55 (d, 3H), 2.65 (s, 3H), 2.89 (s, 3H), 3.00 (s, 3H),3.20-3.27 (m, 2H), 3.27-3.33 (m partially hidden by H2O, 2H), 3.42-3.48(ms, 2H), 3.48-3.56 (m, 2H), 5.48 (q, 1H), 5.54 (s, 1H), 6.83 (dd, 2H),7.04 (dd, 1H), 7.65 (d, 2H), 7.86 (d, 1H).

EXAMPLE 3.088-(1-((3-chloro-4-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-chloro-4-fluoro-N-methylaniline (260 mg, 1.63 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (200 mg, 0.41 mmol, as described in Example 3.03), using ananalogous procedure to that described in Example 3.00, to afford8-(1-((3-chloro-4-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(107 mg, 53.7%) as a white solid. Mass Spectrum: M+H⁺ 488. NMR Spectrum:(DMSOd₆) 1.55 (d, 3H), 2.63 (s, 3H), 2.91 (s, 3H), 3.01 (s, 3H),3.20-3.29 (m, 2H), 3.30-3.37 (m partially hidden by H2O, 2H), 3.42-3.49(ms, 2H), 3.49-3.57 (m, 2H), 5.54 (s, 1H), 5.55 (q, 1H), 6.79 (dd, 1H),6.98 (dd, 1H), 7.23 (dd, 1H), 7.68 (d, 1H), 7.87 (d, 1H).

EXAMPLES 3.09-3.13

For preparation of the compounds of Examples 3.09 to 3.13 (shown inTable I), the appropriate aniline (1.20 mmol) and8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.123 g, 0.3 mmol) were suspended in NMP (1.0 mL) andsealed into a tube. The reaction was purged with argon and heated at 75°C. over a period of 15 hrs. The reaction mixture was purified bypreparative HPLC using a Waters X-Bridge reverse-phase column (C-18, 5microns silica, 19 mm diameter, 100 mm length, flow rate of 40mL/minute) and decreasingly polar mixtures of water (containing 0.2%ammonium carbonate and acetonitrile as eluent. The fractions containingthe desired compound were evaporated to dryness.

TABLE I Product Reagent Mass Yield Ex. Structure Name Product (g) (%)MH+ 3.09

3- chloroaniline 8-(1-(3- chlorophenylamino)ethyl)- N,N-dimethyl-2-morpholino-4-oxo-4H- chromene-6- carboxamide 0.068 49.7 456 3.10

2,3- difluoroaniline 8-(1-(2,3- difluorophenylamino)ethyl)-N,N-dimethyl-2- morpholino-4-oxo-4H- chromene-6- carboxamide0.059 43.0 458 3.11

3,4,5- trifluoroaniline N,N-dimethyl-2- morpholino-4-oxo-8-(1- (3,4,5-trifluorophenylamino) ethyl)-4H-chromene-6- carboxamide 0.074 51.9 4763.12

3-fluoroaniline 8-(1-(3- fluorophenylamino)ethyl)- N,N-dimethyl-2-morpholino-4-oxo-4H- chromene-6- carboxamide 0.094 71.3 440 3.13

2,3,5- trifluoroaniline N,N-dimethyl-2- morpholino-4-oxo-8-(1- (2,3,5-trifluorophenylamino) ethyl)-4H-chromene-6- carboxamide 0.049 74 476Notes Further characterising data for the products is given below.

EXAMPLE 3.09

NMR Spectrum: (CDCl₃) 1.62 (d, 3H), 2.85 (s, 3H), 3.06 (s, 3H),3.42-3.55 (m, 4H), 3.76-3.77 (m, 4H), 4.14 (d, 1H), 4.90-5.00 (m, 1H),5.54 (s, 1H), 6.34 (dd, 1H), 6.45 (dd, 1H), 6.64 (dd, 1H), 7.01 (dd,1H), 7.71 (d, 1H), 8.12 (d, 1H).

EXAMPLE 3.10

NMR Spectrum: (CDCl₃) 1.67 (d, 3H), 2.86 (s, 3H), 3.07 (s, 3H),3.44-3.57 (m, 4H), 3.79-3.88 (m, 4H), 4.38 (bs, 1H), 4.94-5.02 (m, 1H),5.55 (s, 1H), 6.08 (dd, 1H), 6.48 (dd, 1H), 6.74 (dd, 1H), 7.71 (d, 1H),8.12 (d, 1H).

EXAMPLE 3.11

NMR Spectrum: (CDCl₃) 1.56 (d, 3H), 2.91 (s, 3H), 3.07 (s, 3H),3.45-3.59 (m, 4H), 3.78-3.92 (m, 4H), 4.39 (d, 1H), 4.77-4.88 (m, 1H),5.55 (s, 1H), 6.01 (dd, 2H), 7.70 (d, 1H), 8.12 (d, 1H).

EXAMPLE 3.12

NMR Spectrum: (CDCl₃) 1.60 (d, 3H), 2.85 (s, 3H), 3.06 (s, 3H),3.45-3.55 (m, 4H), 3.76-3.87 (m, 4H), 4.22 (d, 1H), 4.89-4.99 (m, 1H),5.55 (s, 1H), 6.13 (ddd, 1H), 6.26 (dd, 1H), 6.36 (ddd, 1H), 7.04 (dd,1H), 7.73 (d, 1H), 8.11 (d, 1H).

EXAMPLE 3.13

NMR Spectrum: (CDCl₃): 1.67 (d, 3H), 2.92 (s, 3H), 3.09 (s, 3H),3.46-3.58 (m, 4H), 3.81-3.90 (m, 4H), 4.53 (bs, 1H), 4.88-4.96 (m, 1H),5.56 (s, 1H), 5.80-5.88 (m, 1H), 6.17-6.27 (m, 1H), 7.71 (d, 1H), 8.13(d, 1H).

A larger batch of this compound was prepared and the enantiomersseparated as follows:

Instrument Kronlab Column Amicon 100 mm Chiralpak IC 20 μm EluentDCM/EtOH/HOAc/TEA 50/50/0.2/0.1 Oven Temperature Ambient Flow 350 ml/minWavelength 254 nm Sample Conc 4.0 g/100 ml in DCM/EtOH 50/50 Injectionvolume 50 mL Run Time 20 min4.2 g of the compound of Example 3.13 was chromatographed in 2injections using the above conditions. Each enantiomer was dissolved inMeOH and added onto an SCX column. The column was flushed with MeOH thenthe product eluted with 7M ammonia in MeOH. The solvents wereevaporated, giving a glass which was slurried with MTBE (75 mL) for 48hrs until it had all turned into a white powdery solid. This wascollected by filtration, washed with MTBE and dried under vacuum at 50°C.First eluted enantiomer: 2.0 g isolated [α]^(D) _(20°): +50° in DCM(enantiomer 1) Example 3.13a.Second eluted enantiomer: 1.9 g isolated [α]^(D) _(20°): −50° in DCM(enantiomer 2) Example 3.13b.

The Second eluting enantiomer (Example 3.13b) was crystallized in DCM,before drying under ambient conditions to yield Form A material. Thisform was determined to be crystalline by XRPD (see FIG. 5) and had thefollowing characteristic X-Ray Powder Diffraction peaks:

Angle 2- Theta (2θ) Intensity % 20.0 100.0 18.0 80.2 14.0 62.0 19.4 51.823.2 29.7 23.8 29.4 10.8 28.4 19.1 28.2 11.2 24.4 27.8 21.8DSC analysis of Form A was also carried out (FIG. 6) and showed thatthis material had a melting point of 156.0° C. (onset).

Form B material was produced by slurrying Form A material in awater/methanol mixture. Approximately 20 mg of the original material wasplaced in a vial with a magnetic flea, and approximately 100 mcL ofmethanol and 2 mL of water added, the vial was then sealed tightly witha cap and left to stir on a magnetic stirrer plate. After 3 days, thesample was removed from the plate, the cap taken off and the slurry leftto dry under ambient conditions before it was analysed by XRPD and DSC.This form (Form B) was determined to be crystalline by XRPD (FIG. 7) andseen to be different to Form A. The form had the followingcharacteristic X-Ray Powder Diffraction peaks:

Angle 2- Theta (2θ) Intensity % 6.2 100.0 7.0 20.4 10.3 10.0 22.4 8.315.9 7.7 20.4 7.5 27.2 6.9 12.4 6.6 18.7 6.3 12.8 6.3

This material had an onset of desolvation at 98.6° C. (onset) (see FIG.8) and thermogravemetirc analysis showed the material to have a massloss consistent with a 1:1 methanol solvate (FIG. 9).

EXAMPLE 3.148-(1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-chloro-5-fluoroaniline (0.074 mL, 0.73 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.09 g, 0.18 mmol)) using an analogous procedure to theone described in Example 3.03 to give8-(1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(0.065 g, 75%) as a white solid. Mass Spectrum: M+H⁺ 474. NMR Spectrum(CDCl3): 1.61 (d, 3H), 2.90 (bs, 3H), 3.09 (bs, 3H), 3.46-3.55 (m, 4H),3.80-3.88 (m, 4H), 4.35 (d, 1H), 4.88-4.96 (m, 1H), 5.56 (s, 1H), 6.03(ddd, 1H), 6.27 (dd, 1H), 6.39 (ddd, 1H), 7.70 (d, 1H), 8.12 (d, 1H).

EXAMPLE 3.158-(1-(2,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2,5-difluoroaniline (0.074 mL, 0.73 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.09 g, 0.18 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give 8-(1-(2,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(0.055 g, 66%) as a white solid. Mass Spectrum: M+H⁺ 458. NMR Spectrum(DMSOd6): 1.57 (d, 3H), 2.72 (bs, 3H), 2.94 (bs, 3H), 3.51-3.64 (m, 4H),3.71-3.78 (m, 4H), 5.02-5.11 (m, 1H), 5.61 (s, 1H), 6.19-1.33 (m, 2H),6.43 (d, 1H), 7.01-7.09 (m, 1H), 7.60 (d, 1H), 7.79 (d, 1H).

EXAMPLE 3.168-(1-(3-fluoro-5-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-fluoro-5-methylaniline (0.091 mL, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.1 g, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-fluoro-5-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(0.053 g, 57%) as a white solid. Mass Spectrum: M+H⁺ 454.

NMR Spectrum (CDCl3): 1.60 (d, 3H), 2.20 (s, 3H), 2.87 (bs, 3H), 3.07(bs, 3H), 3.45-3.54 (m, 4H), 3.79-3.85 (m, 4H), 4.11 (d, 1H), 4.90-4.97(m, 1H), 5.55 (s, 1H), 5.92 (ddd, 1H), 6.12 (s, 1H), 6.20 (d, 1H), 7.73(d, 1H), 8.12 (d, 1H).

EXAMPLE 3.178-(1-(3-cyano-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-amino-5-fluorobenzonitrile (0.111 g, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.1 g, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-cyano-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(0.052 g, 55%) as a white solid. Mass Spectrum: M+H⁺ 465.

NMR Spectrum (CDCl3): 1.62 (d, 3H), 2.92 (bs, 3H), 3.08 (bs, 3H),3.45-3.58 (m, 4H), 3.82-3.91 (m, 4H), 4.61 (d, 1H), 4.88-4.97 (m, 1H),5.57 (s, 1H), 6.36 (ddd, 1H), 6.49 (dd, 1H), 6.65 (ddd, 1H), 7.70 (d,1H), 8.14 (d, 1H).

EXAMPLE 3.188-(1-(3-cyanophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-aminobenzonitrile (96 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.1 g, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-cyanophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(44 mg, 48%) as a white solid. Mass Spectrum: M+H⁺ 447.

NMR Spectrum (CDCl3): 1.63 (d, 3H), 2.88 (bs, 3H), 3.07 (bs, 3H),3.45-3.58 (m, 4H), 3.81-3.88 (m, 4H), 4.35 (d, 1H), 4.91-4.99 (m, 1H),5.57 (s, 1H), 6.64 (s, 1H), 6.70 (dd, 1H), 6.96 (d, 1H), 7.19 (dd, 1H),7.71 (d, 1H), 8.13 (d, 1H).

EXAMPLE 3.198-(1-(2,3-dichlorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2,3-dichloroaniline (0.097 mL, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.1 g, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(2,3-dichlorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(51 mg, 51%) as a white solid. Mass Spectrum: M+H⁺ 490.

NMR Spectrum (CDCl3): 1.69 (d, 3H), 2.85 (s, 3H), 3.07 (s, 3H),3.47-3.56 (m, 4H), 3.79-3.88 (m, 4H), 4.86 (d, 1H), 4.93-5.02 (m, 1H),5.56 (s, 1H), 6.19 (d, 1H), 6.78 (dd, 1H), 6.89 (dd, 1H), 7.66 (d, 1H),8.12 (d, 1H).

EXAMPLE 3.208-(1-(3-ethynylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-ethynylaniline (0.083 mL, 0.73 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (0.09 g, 0.18 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-ethynylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(52 mg, 64%) as a white solid. Mass Spectrum: M+H⁺ 446.

NMR Spectrum (DMSOd6): 1.52 (d, 3H), 2.68 (bs, 3H), 2.93 (bs, 3H),3.50-3.64 (m, 4H), 3.69-3.79 (m, 4H), 3.99 (s, 1H), 4.98-5.06 (m, 1H),5.60 (s, 1H), 6.46-6.54 (m, 2H), 6.57-6.63 (m, 2H), 7.00 (dd, 1H), 7.56(d, 1H), 7.78 (d, 1H).

EXAMPLE 3.218-(1-(5-cyano-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-amino-4-methylbenzonitrile (108 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(5-cyano-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(50 mg, 53%) as a white solid. Mass Spectrum: M+H⁺ 461. NMR Spectrum(DMSOd6): 1.62 (d, 3H), 2.30 (s, 3H), 2.69 (bs, 3H), 2.93 (bs, 3H),3.51-3.64 (m, 4H), 3.70-3.80 (m, 4H), 5.00-5.08 (m, 1H), 5.56 (d, 1H),5.61 (s, 1H), 5.96 (dd, 1H), 6.24 (ddd, 1H), 6.96 (dd, 1H), 7.58 (d,1H), 7.79 (d, 1H).

EXAMPLE 3.228-(1-(5-fluoro-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

5-fluoro-2-methylaniline (102 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(5-fluoro-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(56 mg, 61%) as a white solid. Mass Spectrum: M+H⁺ 454. NMR Spectrum(DMSOd6): 1.60 (d, 3H), 2.20 (s, 3H), 2.69 (bs, 3H), 2.93 (bs, 3H),3.53-3.65 (m, 4H), 3.72-3.80 (m, 4H), 5.08-5.18 (m, 1H), 5.62 (s, 1H),5.73 (d, 1H), 6.52 (d, 1H), 6.92 (dd, 1H), 7.18 (dd, 1H), 7.58 (d, 1H),7.80 (d, 1H).

EXAMPLE 3.238-(1-(3-cyano-5-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-amino-5-methylbenzonitrile (108 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure similar tothe one described in Example 3.03 to give8-(1-(3-cyano-5-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(45 mg, 48%) as a white solid. Mass Spectrum: M+H⁺ 461.

NMR Spectrum (DMSOd6): 1.53 (d, 3H), 2.16 (s, 3H), 2.71 (bs, 3H), 2.94(bs, 3H), 3.51-3.64 (m, 4H), 3.69-3.80 (m, 4H), 5.00-5.10 (m, 1H), 5.61(s, 1H), 6.60 (s, 1H), 6.67 (s, 1H), 6.73 (d, 1H), 6.74 (s, 1H), 7.54(d, 1H), 7.79 (d, 1H).

EXAMPLE 3.248-(1-(5-fluoro-2-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

5-fluoro-2-methoxyaniline (115 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(5-fluoro-2-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(54 mg, 56%) as a white solid. Mass Spectrum: M+H⁺ 470. NMR Spectrum(DMSOd6): 1.58 (d, 3H), 2.71 (bs, 3H), 2.94 (bs, 3H), 3.51-3.64 (m, 4H),3.70-3.78 (m, 4H), 3.82 (s, 3H), 4.97-5.06 (m, 1H), 5.61 (s, 1H), 6.60(s, 1H), 6.67 (s, 1H), 6.73 (d, 1H), 6.74 (s, 1H), 7.54 (d, 1H), 7.79(d, 1H).

EXAMPLE 3.258-(1-(5-cyano-2-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-amino-4-methoxybenzonitrile (121 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(5-cyano-2-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(50 mg, 51%) as a white solid. Mass Spectrum: M+H⁺ 477.

NMR Spectrum (DMSOd6): 1.60 (d, 3H), 2.72 (bs, 3H), 2.94 (bs, 3H),3.51-3.65 (m, 4H), 3.71-3.80 (m, 4H), 3.92 (s, 3H), 5.05-5.13 (m, 1H),5.61 (s, 1H), 5.86 (d, 1H), 6.57 (d, 1H), 6.98 (d, 1H), 7.03 (dd, 1H),7.57 (d, 1H), 7.79 (d, 1H).

EXAMPLE 3.268-(1-(2-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2-chloro-5-fluoroaniline (119 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(2-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(45 mg, 47%) as a white solid. Mass Spectrum: M+H⁺ 474. NMR Spectrum(DMSOd6): 1.62 (d, 3H), 2.74 (bs, 3H), 2.94 (bs, 3H), 3.51-3.65 (m, 4H),3.69-3.79 (m, 4H), 5.06-5.16 (m, 1H), 5.61 (s, 1H), 5.99 (d, 1H), 6.27(dd, 1H), 6.40 (ddd, 1H), 7.30 (dd, 1H), 7.59 (d, 1H), 7.80 (d, 1H).

EXAMPLE 3.27N,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,6-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide

2,3,6-trifluoroaniline (0.086 mL, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to giveN,N-dimethyl-2-morpholino-4-oxo-8-(1-(2,3,6-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide(42 mg, 43%) as a white solid. Mass Spectrum: M+H⁺ 476. NMR Spectrum(DMSOd6): 1.56 (d, 3H), 2.75 (bs, 3H), 2.97 (bs, 3H), 3.50-3.59 (m, 4H),3.69-3.80 (m, 4H), 5.38-5.47 (m, 1H), 5.57 (s, 1H), 6.05 (d, 1H),6.60-6.70 (m, 1H), 6.86-6.96 (m, 1H), 7.73 (d, 1H), 7.76 (d, 1H).

EXAMPLE 3.288-(1-(5-chloro-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

5-chloro-2-methylaniline (0.098 mL, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(5-chloro-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(56 mg, 58%) as a white solid. Mass Spectrum: M+H⁺ 470. NMR Spectrum(DMSOd6): 1.61 (d, 3H), 2.21 (s, 3H), 2.71 (bs, 3H), 2.93 (bs, 3H),3.52-3.65 (m, 4H), 3.70-3.80 (m, 4H), 5.03-5.11 (m, 1H), 5.55 (d, 1H),5.62 (s, 1H), 6.19 (d, 1H), 6.49 (dd, 1H), 6.97 (d, 1H), 7.62 (d, 1H),7.79 (d, 1H).

EXAMPLE 3.298-(1-(3-cyano-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-amino-2-methylbenzonitrile (108 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-cyano-2-methylphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(52 mg, 55%) as a white solid. Mass Spectrum: M+H⁺ 461.

NMR Spectrum (DMSOd6): 1.61 (d, 3H), 2.45 (s, 3H), 2.65 (bs, 3H), 2.92(bs, 3H), 3.50-3.64 (m, 4H), 3.70-3.79 (m, 4H), 5.05-5.14 (m, 1H), 5.61(s, 1H), 5.81 (d, 1H), 6.46 (d, 1H), 6.92 (d, 1H), 7.02 (dd, 1H), 7.56(d, 1H), 7.78 (d, 1H).

EXAMPLE 3.308-(1-(3-fluoro-5-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-fluoro-5-methoxyaniline (115 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-fluoro-5-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(60 mg, 63%) as a white solid. Mass Spectrum: M+H⁺ 470. NMR Spectrum(DMSOd6): 1.50 (d, 3H), 2.74 (bs, 3H), 2.94 (bs, 3H), 3.50-3.63 (m, 4H),3.61 (s, 3H), 3.68-3.79 (m, 4H), 4.94-5.03 (m, 1H), 5.60 (s, 1H), 5.87(dd, 1H), 5.88 (s, 1H), 5.93 (ddd, 1H), 6.63 (d, 1H), 7.57 (d, 1H), 7.79(d, 1H).

EXAMPLE 3.318-(1-(3-fluoro-2-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-fluoro-2-methoxyaniline (115 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-fluoro-2-methoxyphenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(57 mg, 60%) as a white solid. Mass Spectrum: M+H⁺ 470. NMR Spectrum(DMSOd6): 1.59 (d, 3H), 2.70 (bs, 3H), 2.93 (bs, 3H), 3.50-3.63 (m, 4H),3.71-3.79 (m, 4H), 3.83 (s, 3H), 5.02-5.11 (m, 1H), 5.60 (s, 1H), 5.95(d, 1H), 6.11 (d, 1H), 6.39 (dd, 1H), 6.72 (ddd, 1H), 7.59 (d, 1H), 7.78(d, 1H).

EXAMPLE 3.328-(1-(3-chloro-2,6-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-chloro-2,6-difluoroaniline (133 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-chloro-2,6-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(47 mg, 47%) as a white solid. Mass Spectrum: M+H⁺ 492. NMR Spectrum(DMSOd6): 1.56 (d, 3H), 2.74 (bs, 3H), 2.97 (bs, 3H), 3.50-3.58 (m, 4H),3.70-3.80 (m, 4H), 5.37-5.46 (m, 1H), 5.57 (s, 1H), 5.99 (d, 1H), 6.81(ddd, 1H), 6.94 (ddd, 1H), 7.72 (d, 1H), 7.76 (d, 1H).

EXAMPLE 3.338-(1-(3,5-dichlorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3,5-dichloroaniline (132 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3,5-dichlorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(75 mg, 75%) as a white solid. Mass Spectrum: M+H⁺ 490. NMR Spectrum(DMSOd6): 1.52 (d, 3H), 2.75 (bs, 3H), 2.95 (bs, 3H), 3.51-3.63 (m, 4H),3.70-3.78 (m, 4H), 5.01-5.09 (m, 1H), 5.61 (s, 1H), 6.50 (s, 2H), 6.61(s, 1H), 6.91 (d, 1H), 7.54 (d, 1H), 7.81 (d, 1H)

EXAMPLE 3.348-(1-(3-ethynyl-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-ethynyl-5-fluoroaniline (165 mg, 1.22 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(3-ethynyl-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(80 mg, 56%) as a white solid. Mass Spectrum: M+H⁺ 464. NMR Spectrum(DMSOd6): 1.52 (d, 3H), 2.72 (bs, 3H), 2.94 (bs, 3H), 3.50-3.63 (m, 4H),3.69-3.78 (m, 4H), 4.12 (s, 1H), 4.98-5.07 (m, 1H), 5.66 (s, 1H), 6.31(d, 1H), 6.40 (d, 1H), 6.46 (s, 1H), 6.81 (d, 1H), 7.54 (d, 1H), 7.80(d, 1H).

EXAMPLE 3.358-(1-(2-cyano-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2-amino-4-fluorobenzonitrile (111 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-(2-cyano-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(40 mg, 42%) as a white solid. Mass Spectrum: M+H⁺ 465.

NMR Spectrum (DMSOd6): 1.62 (d, 3H), 2.76 (bs, 3H), 2.96 (bs, 3H),3.49-3.64 (m, 4H), 3.67-3.79 (m, 4H), 5.12-5.22 (m, 1H), 5.61 (s, 1H),6.35 (dd, 1H), 6.52 (ddd, 1H), 6.88 (d, 1H), 7.61 (dd, 1H), 7.68 (d,1H), 7.82 (d, 1H).

EXAMPLE 3.368-(1-(2-cyano-3-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2-amino-6-fluorobenzonitrile (111 mg, 0.82 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (100 mg, 0.20 mmol) using an analogous procedure to the onedescribed in Example 3.03.

The crude product was purified by flash chromatography on silica geleluting with 0 to 10% MeOH in DCM. The solvent was evaporated todryness. The resulting oil was crystallised from ethyl acetate to give8-(1-(2-cyano-3-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(45.0 mg, 64.3%) as a white solid. Mass Spectrum: M+H⁺ 465. NMR Spectrum(DMSOd6): 1.61 (d, 3H), 2.74 (bs, 3H), 2.95 (bs, 3H), 3.49-3.62 (m, 4H),3.69-3.77 (m, 4H), 5.17-5.26 (m, 1H), 5.61 (s, 1H), 6.33 (d, 1H), 6.56(dd, 1H), 6.99 (d, 1H), 7.32 (dd, 1H), 7.69 (d, 1H), 7.81 (d, 1H).

EXAMPLE 3.378-(1-((3-chloro-5-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3-chloro-5-fluoro-N-methylaniline (195 mg, 1.22 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (150 mg, 0.31 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give8-(1-((3-chloro-5-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(38 mg, 25%) as a white solid. Mass Spectrum: M+H⁺ 488. NMR Spectrum(DMSOd6): 1.55 (d, 3H), 2.61 (s, 3H), 2.93 (bs, 3H), 3.01 (bs, 3H),3.20-3.27 (m, 2H), 3.33-3.37 (m partially hidden by H2O, 2H), 3.42-3.48(m, 2H), 3.49-3.55 (m, 2H), 5.55 (s, 1H), 5.61 (q, 1H), 6.61 (d, 1H),6.66 (d, 1H), 6.75 (s, 1H), 7.71 (d, 1H), 7.89 (d, 1H).

EXAMPLE 3.38N,N-dimethyl-8-(1-(methyl(3,4,5-trifluorophenyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

3,4,5-trifluoro-N-methylaniline (197 mg, 1.22 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (150 mg, 0.31 mmol) using an analogous procedure to the onedescribed in Example 3.03 to giveN,N-dimethyl-8-(1-(methyl(3,4,5-trifluorophenyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(54.0 mg, 36.1%) as a white solid. Mass Spectrum: M+H⁺ 490.

NMR Spectrum (DMSOd6): 1.54 (d, 3H), 2.62 (s, 3H), 2.91 (bs, 3H), 3.00(bs, 3H), 3.24-3.31 (m partially hidden by H2O, 2H), 3.33-3.42 (mpartially hidden by H2O, 2H), 3.45-3.52 (m, 2H), 3.52-3.60 (m, 2H), 5.55q, 1H), 5.56 (s, 1H), 6.73 (d, 1H), 6.75 (d, 1H), 7.67 (d, 1H), 7.89 (d,1H).

EXAMPLE 3.39N,N-dimethyl-8-(1-(methyl(2,3,5-trifluorophenyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2,3,5-trifluoro-N-methylaniline (197 mg, 1.22 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (150 mg, 0.31 mmol) using an analogous procedure to the onedescribed in Example 3.03 to give N,N-dim ethyl-8-(1-(methyl(2,3,5-trifluorophenyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(12 mg, 8%) as a white solid. Mass Spectrum: M+H⁺ 490.

NMR Spectrum (DMSOd6): 1.63 (d, 3H), 2.67 (s, 3H), 2.92 (bs, 3H), 3.02(bs, 3H), 3.18-3.26 (m, 2H), 3.22-3.37 (m partially hidden by H2O, 2H),3.49-3.56 (m, 2H), 3.56-3.63 (m, 2H), 5.38 (q, 1H), 5.53 (s, 1H),6.68-6.75 (m, 1H), 6.88-6.96 (m, 1H), 7.78 (d, 1H), 7.86 (d, 1H).

EXAMPLE 3.40N,N-dimethyl-8-(1-(methyl(phenyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide

N-methylaniline (153 mg, 1.43 mmol) was reacted with8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidehydrobromide (175 mg, 0.36 mmol) using an analogous procedure to the onedescribed in Example 3.03 to giveN,N-dimethyl-8-(1-(methyl(phenyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxamide(57 mg, 37%) as a white solid. Mass Spectrum: M+H⁺ 436. NMR Spectrum(DMSOd6): 1.57 (d, 3H), 2.65 (s, 3H), 2.90 (bs, 3H), 3.00 (bs, 3H),3.16-3.22 (m, 2H), 3.23-3.31 (m partially hidden by H2O, 2H), 3.36-3.42(m, 2H), 3.42-3.49 (m, 2H), 5.52 (s, 1H), 5.55 (q, 1H), 6.66 (t, 1H),6.84 (d, 2H), 7.19 (t, 2H), 7.67 (d, 1H), 7.87 (d, 1H).

EXAMPLE 3.418-(1-((3-ethynyl-5-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

8-(1-bromoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(185 mg, 0.37 mmol), N,N-diethylaniline (174 μl, 1.10 mmol) and3-ethynyl-5-fluoro-N-methylaniline (60 mg, 0.40 mmol) in DMF (1043 μl)were stirred at 50° C. for 2 days. Purification was done using ananalogous procedure to the one described in Example 3.03 to give8-(1-((3-ethynyl-5-fluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(60 mg, 34%) as a white solid. Mass Spectrum: M+H⁺ 478. NMR Spectrum(DMSOd6): 1.55 (d, 3H), 2.62 (s, 3H), 2.92 (bs, 3H), 3.01 (bs, 3H),3.18-3.27 (m, 2H), 3.28-3.33 (m partially hidden by H2O, 2H), 3.40-3.48(m, 2H), 3.48-3.57 (m, 2H), 4.21 (s, 1H), 5.54 (q, 1H), 5.62 (s, 1H),6.52 (d, 1H), 6.73 (d, 1H), 6.80 (s, 1H), 7.72 (d, 1H), 7.89 (d, 1H).

EXAMPLES 4.01-4.26

0.17 mmole of a previously prepared solution of 2,5-dioxopyrrolidin-1-yl8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylatewas introduced into 26 vials, each containing the appropriate amine(0.51 mmol) for each Example compound (as shown under the column named‘Reagent Name’ in Table II). The resulting solution was stirred at 35°C. for 2 hrs, concentrated to dryness and diluted with DMF (1.5 mL). Thereaction mixture was purified by preparative HPLC using a WatersX-Bridge reverse-phase column (C-18, 5 microns silica, 19 mm diameter,100 mm length, flow rate of 40 mL/minute) and decreasingly polarmixtures of water (containing 0.2% ammonium carbonate) and acetonitrileas eluent. The fractions containing the desired compound were evaporatedto dryness to afford the desired product.

TABLE II Product Mass Yield EX. Structure Reagent Name Product (g) (%)MH+ 4.01

diethylamine 8-(1-(3,5- difluorophenylamino) ethyl)-N,N-diethyl-2-morpholino-4-oxo- 4H-chromene-6- carboxamide 46 56.2 486 4.02

pyrrolidine 8-(1-(3,5- difluorophenylamino) ethyl)-2- morpholino-6-(pyrrolidine-1- carbonyl)-4H- chromen-4-one 54 65.1 484 4.03

1-methyl- piperazine 8-(1-(3,5- difluorophenylamino) ethyl)-6-(4-methylpiperazine-1- carbonyl)-2- morpholino-4H- chromen-4-one 60 68.3513 4.04

tert-butyl piperazine-1- carboxylate (2 steps) 8-(1-(3,5-difluorophenylamino) ethyl)-2- morpholino-6- (piperazine-1-carbonyl)-4H- chromen-4-one 39 46.0 499 4.05

thiomorpholine 8-(1-(3,5- difluorophenylamino) ethyl)-2- morpholino-6-(thiomorpholine-4- carbonyl)-4H- chromen-4-one 52 58.6 516 4.06

azepane 6-(azepane-1- carbonyl)-8-(1-(3,5- difluorophenylamino)ethyl)-2- morpholino-4H- chromen-4-one 56 64.1 512 4.07

2-(methylamino) ethanol 8-(1-(3,5- difluorophenylamino) ethyl)-N-(2-hydroxyethyl)-N- methyl-2- morpholino-4-oxo- 4H-chromene-6- carboxamide52 62.5 488 4.08

azetidine 6-(azetidine-1- carbonyl)-8-(1-(3,5- difluorophenylamino)ethyl)-2- morpholino-4H- chromen-4-one 48 59.8 470 4.09

piperidin-4-ol 8-(1-(3,5- difluorophenylamino) ethyl)-6-(4-hydroxypiperidine-1- carbonyl)-2- morpholino-4H- chromen-4-one 55 63.3514 4.10

piperidine 8-(1-(3,5- difluorophenylamino) ethyl)-2- morpholino-6-(piperidine-1- carbonyl)-4H- chromen-4-one 51 60.3 498 4.11

N-methyl- ethanamine 8-(1-(3,5- difluorophenylamino) ethyl)-N-ethyl-N-methyl-2- morpholino-4-oxo- 4H-chromene-6- carboxamide 51 63.9 472 4.12

azetidin-3-ol hydrochloride 8-(1-(3,5- difluorophenylamino) ethyl)-6-(3-hydroxyazetidine-1- carbonyl)-2- morpholino-4H- chromen-4-one 50 60.9486 4.13

3-fluoroazetidine hydrochloride 8-(1-(3,5- difluorophenylamino)ethyl)-6-(3- fluoroazetidine-1- carbonyl)-2- morpholino-4H-chromen-4-one 49 58.9 488 4.14

2-aminoethanol 8-(1-(3,5- difluorophenylamino) ethyl)-N-(2-hydroxyethyl)-2- morpholino-4-oxo- 4H-chromene-6- carboxamide 49 61.0474 4.15

2- methoxyethan- amine 8-(1-(3,5- difluorophenylamino) ethyl)-N-(2-methoxyethyl)-2- morpholino-4-oxo- 4H-chromene-6- carboxamide 53 63.5488 4.16

propan-1-amine 8-(1-(3,5- difluorophenylamino) ethyl)-2-morpholino-4-oxo- N-propyl-4H- chromene-6- carboxamide 44 54 472 4.17

ethanamine hydrochloride 8-(1-(3,5- difluorophenylamino)ethyl)-N-ethyl-2- morpholino-4-oxo- 4H-chromene-6- carboxamide 33 42 4584.18

2- fluoroethanamine hydrochloride 8-(1-(3,5- difluorophenylamino)ethyl)-N-(2- fluoroethyl)-2- morpholino-4-oxo- 4H-chromene-6-carboxamide 43 53.3 476 4.19

3-methoxy- propan-1-amine 8-(1-(3,5- difluorophenylamino) ethyl)-N-(3-methoxypropyl)-2- morpholino-4-oxo- 4H-chromene-6- carboxamide 40 46.5502 4.20

(R)-pyrrolidin-2- ylmethanol 8-(1-(3,5- difluorophenylamino)ethyl)-6-((R)-2- (hydroxymethyl) pyrrolidine-1-carbonyl)-2-morpholino-4H- chromen-4-one 53 60.7 514 4.21

(S)-pyrrolidin-2- ylmethanol 8-(1-(3,5- difluorophenylamino)ethyl)-6-((S)-2- (hydroxymethyl) pyrrolidine-1-carbonyl)-2-morpholino-4H- chromen-4-one 52 59.3 514 4.22

methanamine 8-(1-(3,5- difluorophenylamino) ethyl)-N-methyl-2-morpholino-4-oxo- 4H-chromene-6- carboxamide 22 29.5 444 4.23

cyclopropyl- methanamine N- (cyclopropylmethyl)- 8-(1-(3,5-difluorophenylamino) ethyl)-2- morpholino-4-oxo- 4H-chromene-6-carboxamide 53 63.8 484 4.24

2-methoxy-N- methyl- ethanamine 8-(1-(3,5- difluorophenylamino)ethyl)-N-(2- methoxyethyl)-N- methyl-2- morpholino-4-oxo- 4H-chromene-6-carboxamide 53 62.4 502 4.25

1,4-oxazepane hydrochloride 8-(1-(3,5- difluorophenylamino) ethyl)-2-morpholino-6-(1,4- oxazepane-4- carbonyl)-4H- chromen-4-one 49 56.3 5144.26

4-methoxy- piperidine 8-(1-(3,5- difluorophenylamino) ethyl)-6-(4-methoxypiperidine- 1-carbonyl)-2- morpholino-4H- chromen-4-one 58 64.9528Notes Further characterising data for the products is given below.

EXAMPLE 4.01

NMR Spectrum: (DMSOd₆) at 353° K: 1.01 (t, 6H), 1.56 (d, 3H), 3.25 (bs,4H), 3.52-3.65 (m, 4H), 3.73-3.81 (m, 4H), 4.98-5.06 (m, 1H), 5.55 (s,1H), 6.11-6.22 (m, 3H), 6.72 (d, 1H), 7.54 (d, 1H), 7.78 (d, 1H).

EXAMPLE 4.02

NMR Spectrum: (DMSOd₆) at 323° K: 1.52 (d, 3H), 1.66-1.78 (m, 2H),1.78-1.91 (m, 2H), 3.07-3.23 (m, 2H), 3.37-3.49 (m partially hidden byH2O, 2H), 3.51-3.64 (m, 4H), 3.69-3.81 (m, 4H), 4.98-5.08 (m, 1H), 5.61(s, 1H), 6.16 (dd, 2H), 6.24 (ddd, 1H), 6.97 (d, 1H), 7.67 (d, 1H), 7.93(d, 1H).

EXAMPLE 4.03

NMR Spectrum: (DMSOd₆) 1.52 (d, 3H), 1.93 (bs, 1H), 2.08 (bs, 1H), 2.14(s, 3H), 2.41 (bs, 2H), 3.05 (bs, 2H), 3.50-3.64 (m, 5H), 3.68 (bs, 1H),3.72-3.80 (ms, 4H), 4.98-5.06 (m, 1H), 5.62 (s, 1H), 6.15 (dd, 2H), 6.25(ddd, 1H), 6.98 (d, 1H), 7.46 (d, 1H), 7.78 (d, 1H).

EXAMPLE 4.04

NMR Spectrum: (DMSOd₆) 1.52 (d, 3H), 2.43 (bs, 2H), 2.69 (bs, 2H), 3.00(bs, 2H), 3.46 (bs partially hidden by H2O, 2H), 3.50-3.65 (m, 5H),3.70-3.79 (m, 4H), 4.97-5.05 (m, 1H), 5.61 (s, 1H), 6.14 (dd, 2H), 6.24(ddd, 1H), 6.98 (d, 1H), 7.47 (d, 1H), 7.78 (d, 1H).

The removal of the tert-butyl-carboxylate protecting group was done asfollows: The crude solution was washed with a 10% solution of citricacid (pH-4), water, dried over magnesium sulfate and concentrated. HCl(4N in dioxane; 1066 μl, 4.27 mmol) was added and the mixture wasstirred at room temperature for 30 minutes. The volatils were removedunder vacuum and the residue dissolved into a 10% methanolic ammonia inDCM (5 mL). The insolubles were removed by filtration, the filtrate wasconcentrated and the crude product was purified by preparative HPLC asdescribed above.

EXAMPLE 4.05

NMR Spectrum: (DMSOd₆) 1.52 (d, 3H), 2.32 (bs, 2H), 2.66 (bs, 2H), 3.45(bs partially hidden by H2O, 1H), 3.50-3.64 (m, 5H), 3.68 (bs, 1H),3.70-3.81 (m, 4H), 3.96 (bs, 1H), 4.96-5.06 (m, 1H), 5.61 (s, 1H), 6.14(dd, 2H), 6.25 (ddd, 1H), 6.96 (d, 1H), 7.51 (d, 1H), 7.80 (d, 1H).

EXAMPLE 4.06

NMR Spectrum: (DMSOd₆) 1.11-1.22 (m, 1H), 1.22-1.32 (m, 1H), 1.32-1.43(m, 2H), 1.44-1.55 (m, 2H), 1.52 (d, 3H), 1.62-1.73 (m, 2H), 3.03-3.12(m, 2H), 3.37-3.45 (m partially hidden by H2O, 1H), 3.51-3.70 (m, 5H),3.71-3.79 (m, 4H), 4.97-5.06 (m, 1H), 5.61 (s, 1H), 6.14 (dd, 2H), 6.23(ddd, 1H), 6.98 (d, 1H), 7.45 (d, 1H), 7.73 (d, 1H).

EXAMPLE 4.07

NMR Spectrum: (DMSOd₆) at 323° K: 1.54 (d, 3H), 2.90 (bs, 3H), 3.20 (bspartially hidden by H2O, 2H), 3.47 (bs, 2H), 3.51-3.64 (m, 4H),3.71-3.79 (m, 4H), 4.61 (bs, 1H), 4.97-5.05 (m, 1H), 5.58 (s, 1H), 6.15(dd, 2H), 6.20 (ddd, 1H), 6.83 (d, 1H), 7.58 (d, 1H), 7.84 (d, 1H).

EXAMPLE 4.08

NMR Spectrum: (DMSOd₆) 1.51 (d, 3H), 2.17-2.28 (m, 2H), 3.49-3.64 (m,4H), 3.70-3.79 (m, 4H), 3.97-4.09 (m, 2H), 4.09-4.20 (m, 2H), 4.97-5.06(m, 1H), 5.62 (s, 1H), 6.15 (dd, 2H), 6.23 (ddd, 1H), 7.04 (d, 1H), 7.79(d, 1H), 8.03 (d, 1H).

EXAMPLE 4.09

NMR Spectrum: (DMSOd₆) at 323° K: 1.27 (bs, 2H), 1.54 (d, 3H), 1.64 (bs,2H), 3.04 (bs, 2H), 3.49 (bs, 1H), 3.52-3.62 (m, 4H), 3.63 (bs, 1H),3.66-3.74 (bs, 1H), 3.72-3.80 (m, 4H), 4.66 (d, 1H), 4.97-5.05 (m, 1H),5.58 (s, 1H), 6.15 (dd, 2H), 6.20 (ddd, 1H), 6.86 (d, 1H), 7.53 (d, 1H),7.80 (d, 1H).

EXAMPLE 4.10

NMR Spectrum: (DMSOd₆) at 323° K: 1.39 (bs, 4H), 1.54 (d, 3H), 1.54-4.64(m, 2H), 3.31 (bs partially hidden by H2O, 4H), 3.49-3.66 (m, 4H),3.68-3.84 (m, 4H), 4.96-5.07 (m, 1H), 5.58 (s, 1H), 6.14 (dd, 2H), 6.20(ddd, 1H), 6.86 (d, 1H), 7.52 (d, 1H), 7.79 (d, 1H).

EXAMPLE 4.11

NMR Spectrum: (DMSOd₆) at 323° K: 0.96 (bs, 3H), 1.54 (d, 3H), 2.85 (bs,3H), 3.23 (bs partially hidden by H2O, 2H), 3.51-3.65 (m, 4H), 3.75-3.82(m, 4H), 4.97-5.06 (m, 1H), 5.58 (s, 1H), 6.15 (dd, 2H), 6.20 (ddd, 1H),6.86 (d, 1H), 7.54 (s, 1H), 7.79 (d, 1H).

EXAMPLE 4.12

NMR Spectrum: (DMSOd₆) at 323° K: 1.53 (d, 3H), 3.51-3.64 (m, 4H),3.71-3.80 (m, 4H), 3.84 (bs, 2H), 4.19-4.32 (m, 2H), 4.43-4.52 (m, 1H),4.97-5.06 (m, 1H), 5.59 (s, 1H), 5.63 (d, 1H), 6.16 (dd, 2H), 6.20 (ddd,1H), 6.92 (d, 1H), 7.80 (s, 1H), 8.05 (d, 1H).

EXAMPLE 4.13

NMR Spectrum: (DMSOd₆) 323° K: 1.53 (d, 3H), 3.52-3.66 (m, 4H),3.71-3.82 (m, 4H), 4.04-4.24 (m, 2H), 4.29-4.51 (m, 2H), 4.98-5.09 (m,1H), 5.39 (ddddd, 1H), 5.59 (s, 1H), 6.17 (dd, 2H), 6.20 (ddd, 1H), 6.91(d, 1H), 7.81 (s, 1H), 8.06 (d, 1H).

EXAMPLE 4.14

NMR Spectrum: (DMSOd₆) 1.53 (d, 3H), 3.27-3.33 (m, 2H), 3.47-3.52 (m,2H), 3.52-3.63 (m, 4H), 3.70-3.79 (m, 4H), 4.73 (t, 1H), 4.96-5.04 (m,1H), 5.62 (s, 1H), 6.15 (dd, 2H), 6.22 (ddd, 1H), 7.03 (d, 1H), 8.08 (s,1H), 8.38 (d, 1H), 8.69 (t, 1H).

EXAMPLE 4.15

NMR Spectrum: (DMSOd₆) 1.52 (d, 3H), 3.26 (s, 3H), 3.35-3.42 (mpartially hidden by H2O, 2H), 3.42-3.48 (m, 2H), 3.50-3.64 (m, 4H),3.70-3.79 (m, 4H), 4.96-5.05 (m, 1H), 5.62 (s, 1H), 6.14 (dd, 2H), 6.22(ddd, 1H), 7.03 (d, 1H), 8.07 (s, 1H), 8.37 (d, 1H), 8.78 (t, 1H).

EXAMPLE 4.16

NMR Spectrum: (DMSOd₆) 0.88 (t, 3H), 1.47-1.58 (m, 2H), 1.52 (d, 3H),3.13-3.24 (m, 2H), 3.50-3.64 (m, 4H), 3.70-3.79 (m, 4H), 4.96-5.05 (m,1H), 5.62 (s, 1H), 6.14 (dd, 2H), 6.22 (ddd, 1H), 7.03 (d, 1H), 8.07 (s,1H), 8.36 (d, 1H), 8.73 (t, 1H).

EXAMPLE 4.17

NMR Spectrum: (DMSOd₆) 1.11 (t, 3H), 1.51 (d, 3H), 3.21-3.30 (m, 2H),3.50-3.64 (m, 4H), 3.69-3.80 (m, 4H), 4.96-5.04 (m, 1H), 5.62 (s, 1H),6.14 (dd, 2H), 6.22 (ddd, 1H), 7.03 (d, 1H), 8.07 (s, 1H), 8.35 (d, 1H),8.74 (t, 1H).

EXAMPLE 4.18

NMR Spectrum: (DMSOd₆) 1.52 (d, 3H), 3.48-3.64 (m, 6H), 3.70-3.78 (m,4H), 4.53 (dt, 2H), 4.96-5.04 (m, 1H), 5.62 (s, 1H), 6.14 (dd, 2H), 6.22(ddd, 1H), 7.03 (d, 1H), 8.09 (d, 1H), 8.39 (d, 1H), 8.96 (t, 1H).

EXAMPLE 4.19

NMR Spectrum: (DMSOd₆) 1.52 (d, 3H), 1.70-1.79 (m, 2H), 3.23 (s, 3H),3.24-3.30 (m, 2H), 3.36-3.42 (m partially hidden by H2O, 2H), 3.50-3.63(m, 4H), 3.71-3.78 (m, 4H), 4.96-5.04 (m, 1H), 5.62 (s, 1H), 6.14 (dd,2H), 6.22 (ddd, 1H), 7.03 (d, 1H), 8.06 (d, 1H), 8.35 (d, 1H), 8.73 (t,1H).

EXAMPLE 4.20

NMR Spectrum: (DMSOd₆) 1.54 (d, 3H), 1.56-1.98 (m, 4H), 2.75-3.47 (m,2H), 3.48-3.65 (m, 6H), 3.68-3.82 (m, 4H), 4.11 (bs, 1H), 4.76-4.86 (m,1H), 4.95-5.07 (m, 1H), 5.61 (s, 1H), 6.15 (dd, 2H), 6.23 (ddd, 1H),6.93-7.02 (m, 1H) 7.64 (s, 0.5H), 7.68 (s, 0.5H), 7.95 (s, 1H).

EXAMPLE 4.21

NMR Spectrum: (DMSOd₆) 1.54 (d, 3H), 1.56-1.98 (m, 4H), 2.75-3.47 (m,2H), 3.48-3.65 (m, 6H), 3.68-3.82 (m, 4H), 4.11 (bs, 1H), 4.76-4.86 (m,1H), 4.95-5.07 (m, 1H), 5.61 (s, 1H), 6.15 (dd, 2H), 6.23 (ddd, 1H),6.93-7.02 (m, 1H) 7.64 (s, 0.5H), 7.68 (s, 0.5H), 7.95 (s, 1H).

EXAMPLE 4.22

NMR Spectrum: (DMSOd₆) 1.51 (d, 3H), 1.75 (d, 3H), 3.51-3.62 (m, 4H),3.71-3.78 (m, 4H), 4.96-5.04 (m, 1H), 5.61 (s, 1H), 6.14 (dd, 2H), 6.22(ddd, 1H), 7.03 (d, 1H), 8.07 (d, 1H), 8.33 (d, 1H), 8.67 (q, 1H).

EXAMPLE 4.23

NMR Spectrum: (DMSOd₆) 0.18-0.27 (m, 2H), 0.39-0.48 (m, 2H), 0.98-1.08(m, 1H), 1.51 (d, 3H), 3.08-3.16 (m, 2H), 3.51-3.64 (m, 4H), 3.71-3.80(m, 4H), 4.97-5.06 (m, 1H), 5.63 (s, 1H), 6.15 (dd, 2H), 6.23 (ddd, 1H),7.05 (d, 1H), 8.09 (d, 1H), 8.38 (d, 1H), 8.86 (t, 1H).

EXAMPLE 4.24

NMR Spectrum: (DMSOd₆) at 323° K: 1.52 (d, 3H), 2.89 (s, 3H), 3.02-3.50(m partially hidden by H2O, 7H), 3.50-3.68 (m, 4H), 3.71-3.80 (m, 4H),5.01 (bs, 1H), 5.63 (s, 1H), 6.14 (dd, 2H), 6.23 (ddd, 1H), 6.98 (d,1H), 7.52 (d, 1H), 7.78 (d, 1H).

EXAMPLE 4.25

NMR Spectrum: (DMSOd₆) at 323° K: 1.43 (bs, 2H), 1.55 (d, 3H), 1.87 (bs,1H), 3.27-3.89 (m, 15H), 4.97-5.06 (m, 1H), 5.59 (s, 1H), 6.15 (dd, 2H),6.21 (ddd, 1H), 6.87 (d, 1H), 7.54 (d, 1H), 7.80 (d, 1H).

EXAMPLE 4.26

NMR Spectrum: (DMSOd₆) at 323° K: 1.34 (bs, 2H), 1.51 (d, 3H), 1.72 (bs,2H), 3.08 (bs, 2H), 3.25 (s, 3H), 3.36-3.44 (m, 1H), (m, 4H), 3.67 (bs,2H), 3.69-3.82 (m, 4H), 4.97-5.07 (m, 1H), 5.58 (s, 1H), 6.15 (dd, 2H),6.21 (ddd, 1H), 6.86 (d, 1H), 7.52 (d, 1H), 7.80 (d, 1H).

The 2,5-dioxopyrrolidin-1-yl8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylateused as starting material for the preparation of compounds of Examples4.01-4.26 was made as follows:—

TSTU (2.098 g, 6.97 mmol) was added at room temperature to8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (made using aprocedure similar to the one described for the synthesis of8-(1-(3-chloro-2-fluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid in Example 2.00;2.5 g, 3.49 mmol) and DIPEA (1.214 mL, 6.97 mmol) in DCM (25 mL). Theresulting solution was stirred for 2 hrs. This solution of intermediatewas used as such for the next step.

EXAMPLE 5.0 8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (2.134 g, 7.09 mmol), was added in one portion to astirred solution of8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (2.1 g, 4.73 mmol) and N-ethyl-N-isopropylpropan-2-amine (1.646 mL,9.45 mmol) in DCM (20 mL) at RT and stirred at RT for 90 mins.Dimethylamine (4.73 mL, 9.45 mmol) was then added and the reactionmixture was stirred at RT for 30 min. Water and DCM were added, theorganic phase was washed with brine, dried over magnesium sulfate andconcentrated. The crude product was purified by flash chromatography onsilica gel eluting with DCM/MeCN (1:1) then 0 to 10% MeOH in DCM. Thesolvent was evaporated to dryness to afford a foam which crystallisedfrom ethyl acetate (10 mL). Ether (10 mL) was added to complete thecrystallization and the white solid was collected by filtration anddried to give 8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(1.65 g, 74%). Mass Spectrum: M+H⁺ 472. NMR Spectrum (DMSOd6): 1.55 (d,3H), 2.63 (s, 3H), 2.92 (bs, 3H), 3.01 (bs, 3H), 3.21-3.29 (m, 2H),3.31-3.39 (m partially hidden by H2O, 2H), 3.41-3.49 (m, 2H), 3.49-3.57(m, 2H), 5.55 (s, 1H), 5.58 (q, 1H), 6.40 (t, 1H), 6.53 (d, 2H), 7.70(d, 1H), 7.89 (d, 1H).

This racemic compound was resolved by chiral preparative HPLC using thefollowing conditions:

Column Chiralpak IA; 21 × 250 mm, 5μ Eluent CO2/MeOH 75:25 OvenTemperature 40° C. Flow 60 mL/min Wavelength 220 nm Sample Conc 50 mg/mLin MeOH Injection 50 mg

1.5 g of racemic was separated using the above conditions to give:

First eluting enantiomer 0.7 g (ee>98%) (Example 5.0a) [α]^(D) _(20°):+5° in MeCN

Second eluting enantiomer 0.7 g (ee>98%) (Example 5.0b) [α]^(D) _(20°):−5° in MeCN

The8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid used as starting material was made as follows:—

Potassium iodide (1.521 g, 9.16 mmol) was added to a suspension ofmethyl 8-(1-bromoethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate(3.3 g, 8.33 mmol, as described in Example 2.00) and3,5-difluoro-N-methylaniline (3.58 g, 24.99 mmol) in CHCl3 (16 mL) andMeOH (4 mL). The mixture was stirred at RT over the weekend. Thereaction mixture was concentrated to dryness and the resulting dark oilwas triturated with diethyl ether to give a solid which was collected byfiltration. This solid was suspended in water and pH was adjusted to 6-7with 2N NaOH. The crude product was filtered and washed with ether,dried and purified by flash chromatography on silica gel eluting with 0to 10% MeOH in DCM. The solvent was evaporated to dryness to affordmethyl8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate(3.1 g, 81%) as a white solid. Mass Spectrum: M+H⁺ 459.

2N NaOH (6.54 mL, 13.1 mmol), was added dropwise to a stirred suspensionof methyl8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylate(3 g, 6.54 mmol), in THF (30 mL)/MeOH (30 mL). The resulting solutionwas stirred at RT overnight. The reaction mixture was diluted withwater, the pH was adjusted to 3 with a 2M aq. sol. HCl. The solventswere removed and the white precipitate was collected by filtration,washed with water and dried then washed with ethyl acetate and ether toafford8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (2.5 g, 86%) which was used without further purification. MassSpectrum: M+H⁺ 445.

The compound of Example 5.0a could also be made using the followingalternative method:

Lithium bis(trimethylsilyl)amide (1N in THF) (26.2 mL, 26.23 mmol) wasadded to a stirred solution of8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(6 g, 13.12 mmol, prepared as described in Example 3.06b; [α]^(D)₂₀=−122.6° in EtOH) dissolved in anhydrous THF (60 mL) at −60° C. underargon. The light red solution was allowed to warm to −10° C. over aperiod of 15 minutes then cooled down to −60° C. before addition ofdimethyl sulfate (2.482 mL, 26.23 mmol). The resulting light yellowsolution was allowed to warm to 0° C. and stirred for 15 minutes. Thereaction mixture was cooled back to −10° C. prior to addition of a sat.aq. sol. of NH₄Cl (30 mL) followed by extraction with DCM. The crudeproduct (7 g) was purified by flash chromatography on silica gel elutingwith 0 to 15% EtOH in DCM/ethyl acetate (1/1) then 15% EtOH in DCM. Thesolvent was evaporated to dryness to afford (4.2 g, 8.91 mmol, 67.9%) asan off-white foam.

EXAMPLE 5.018-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one

2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (113 mg, 0.35 mmol) was added to a stirred solution of8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (130 mg, 0.29 mmol), 4-methylmorpholine (0.080 mL, 0.73 mmol) andpiperidin-4-ol (36 mg, 0.35 mmol) dissolved in NMP (1.2 mL). Theresulting solution was stirred at 23° C. for 2 hrs. The reaction mixturewas purified by preparative HPLC using a Waters SunFire system. Thefractions containing the desired compound were evaporated, trituratedwith ether and dried to afford8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(90 mg, 58%) as a white solid. Mass Spectrum: M+H⁺ 528. NMR Spectrum(DMSOd6 at 323° K): 1.38 (bs, 2H), 1.57 (d, 3H), 1.75 (bs, 2H), 2.66 (s,3H), 3.23-3.30 (m, 2H), 3.30-3.38 (m, 2H), 3.61 (m, 4H), 3.65 (bs, 4H),3.71-3.79 (m, 1H), 4.66 (d, 1H), 5.52 (s, 1H), 5.54 (q, 1H), 6.36 (t,1H), 6.50 (d, 2H), 7.62 (d, 1H), 7.87 (d, 1H).

EXAMPLE 5.028-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(3-hydroxypyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one

This compound was prepared using an analogous procedure to the onedescribed in Example 5.01. Pyrrolidin-3-ol (0.028 mL, 0.35 mmol) wasused in place of piperidin-4-ol to give8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(3-hydroxypyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(80 mg, 53%) as a white solid. Mass Spectrum: M+H⁺ 514. NMR Spectrum(DMSOd6 at 323° K): 1.57 (d, 3H), 1.82 (bs, 1H), 1.96 (bs, 1H), 2.63 (s,1.5H), 2.65 (s, 1.5H), 3.22-3.62 (m, 12H), 4.25 (bs, 0.5H), 4.33 (bs,0.5H), 4.86 (bs, 0.5H), 4.92 (bs, 0.5H), 5.51 (s, 1H), 5.56 (q, 1H),6.36 (t, 1H), 6.51 (d, 2H), 7.76 (d, 0.5H), 7.78 (bs, 0.5H), 8.02 (bs,1H).

EXAMPLE 5.03

8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-((R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one

This compound was prepared using a procedure similar to the onedescribed in Example 5.01. (R)-pyrrolidin-2-ylmethanol (0.035 mL, 0.35mmol) was used in place of piperidin-4-ol to give8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-((R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(81 mg, 53%) as a white solid. Mass Spectrum: M+H⁺ 528. NMR Spectrum(DMSOd6 at 323° K): 1.57 (d, 3H), 1.72 (bs, 1H), 1.83-2.01 (m, 3H), 2.61(s, 1.5H), 2.65 (s, 1.5H), 3.22-3.68 (m, 12H), 4.17 (bs, 1H), 4.68 (bs,1H), 5.51 (s, 0.5H), 5.52 (s, 0.5H), 5.53-5.60 (m, 1H), 6.36 (t, 1H),6.47-6.55 (m, 2H), 6.74 (s, 0.5H), 6.78 (s, 0.5H), 8.01 (bs, 1H).

EXAMPLE 6.0 8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-one

3,5-difluoro-N-methylaniline (222 mg, 1.55 mmol),8-(1-bromoethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-onehydrobromide (200 mg, 0.39 mmol) and potassium iodide (64.3 mg, 0.39mmol) in CHCl3 (0.8 mL) and MeOH (0.2 mL) were stirred at 20° C. for 25hrs. The reaction mixture was concentrated to dryness, diluted with DCM(30 mL), washed with water, brine, dried over magnesium sulfate andconcentrated to afford the crude product. Purification was done by flashchromatography on silica gel eluting with 2 to 4% MeOH in DCM. Thesolvent was evaporated to dryness to afford a foam which was dissolvedin acetonitrile-water and concentrated under vacuum to give8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-one (115 mg,60%) as a white solid. Mass Spectrum: M+H⁺ 498. NMR Spectrum (DMSOd6):1.56 (d, 3H), 1.78-1.93 (m, 4H), 2.62 (s, 3H), 3.23-3.29 (m, 2H),3.35-3.56 (m, 10H), 5.55 (s, 1H), 5.59 (q, 1H), 6.40 (t, 1H), 6.53 (d,2H), 7.79 (d, 1H), 8.01 (d, 1H).

The8-(1-bromoethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-onehydrobromide used as starting material was made as follows:—

2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (3.34 g, 11.09 mmol) at 25° C. was added portionwiseto 8-(1-hydroxyethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid(1.77 g, 5.54 mmol) and N-ethyl-N-isopropylpropan-2-amine (2.028 mL,11.64 mmol) suspended in DCM (15 mL) under nitrogen. The resultingmixture was stirred at 25° C. for 5 hrs. Pyrrolidine (1.388 mL, 16.63mmol) was then added to the mixture and the resulting mixture wasstirred at 25° C. overnight. The mixture was poured onto a silica gelcolumn and purified by flash chromatography eluting with 2 to 7%methanolic ammonia (7 N) in DCM. The solvent was evaporated to dryness,the residue was triturated in ethylacetate (10 mL), collected byfiltration and dried to afford8-(1-hydroxyethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-one(1.66 g, 80%) as a beige solid. Mass Spectrum: M+H⁺ 373.

A solution of tribromophosphine (0.491 mL, 5.22 mmol) in1,2-dichloroethane (4 mL) at 10° C., was added dropwise to8-(1-hydroxyethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-one(1.62 g, 4.35 mmol) suspended in 1,2-dichloroethane (18 mL) undernitrogen. The resulting suspension was stirred at 50° C. for 1 hour. Thereaction mixture was allowed to cool to RT under stirring and dilutedwith diethyl ether (18 mL). The precipitate was collected by filtration,washed with diethyl ether and dried to a constant weight to afford8-(1-bromoethyl)-2-morpholino-6-(pyrrolidine-1-carbonyl)-4H-chromen-4-one(2.45 g, 100%) hydrobromide as a white solid, which was used withoutfurther purification. NMR Spectrum (DMSOd6): 1.79-1.93 (m, 4H), 2.11 (d,3H), 3.37-3.44 (m, 2H), 3.46-3.53 (m, 2H), 3.57-3.70 (m, 4H), 3.73-3.80(m, 4H), 5.66 (s, 1H), 5.92 (q, 1H), 7.99-8.03 (m, 2H).

EXAMPLE 7.08-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide

TBTU (108 mg, 0.34 mmol) was added in one portion to a stirred solutionof8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (125 mg, 0.28 mmol), N-ethyl-N-isopropylpropan-2-amine (0.103 mL,0.59 mmol) and 2-(methylamino)ethanol (0.027 mL, 0.34 mmol) in DMF (1mL). The resulting solution was stirred at RT for 2 hrs. The reactionmixture was filtered and purified by preparative HPLC using areverse-phase column (C-18, 5 microns silica, 19 mm diameter, 100 mmlength, flow rate of 40 mL/minute) and decreasingly polar mixtures ofwater (containing 0.2% ammonium carbonate) and acetonitrile as eluent.The fractions containing the desired compound were evaporated to drynessto afford8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(90 mg, 64%) as a off-white solid. Mass Spectrum: M+H⁺502. NMR Spectrum(DMSOd6 at 323° K): 1.58 (d, 3H), 2.65 (s, 3H), 3.01 (s, 3H), 3.23-3.40(m, 5H), 3.4-3.71 (m, 7H), 4.74 (bs, 1H), 5.52 (s, 1H), 5.57 (q, 1H),6.38 (t, 1H), 6.53 (d, 2H), 7.74 (bs, 1H), 7.93 (s, 1H).

EXAMPLE 7.0A

8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(enantiomer 1)

Tetrabutylammonium fluoride (0.568 mL, 0.57 mmol) was added to a stirredsolution ofN-(2-(tert-butyldiphenylsilyloxy)ethyl)-8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamideenantiomer 1 (210 mg, 0.28 mmol) dissolved in THF (2 mL) at RT undernitrogen and the resulting solution stirred for 16 hrs. The mixture wasevaporated to dryness, diluted with DCM and washed with water, brine,dried over magnesium sulfate, filtered and concentrated. The crudeproduct was purified by flash chromatography on silica gel eluting with5 to 7% MeOH in DCM. The solvent was evaporated to dryness, the gum wastriturated in ether/pentane, the solid was collected by filtration anddried to give8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamideenantiomer 1 (79 mg, 56%) as a white solid. Mass Spectrum: M+H⁺ 502.[α]^(D) _(20°): −9° in MeCN.

TheN-(2-(tert-butyldiphenylsilyloxy)ethyl)-8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide enantiomer1 used as starting material was made as follows:—

To a solution of 2-(methylamino)ethanol (2.14 mL, 26.6 mmol) in DCM (60mL) was added triethylamine (4.1 mL, 29.3mmol),N,N-dimethylpyridin-4-amine (1.63 g, 13.3 mmol) andtert-butylchlorodiphenylsilane (7.6 mL, 29.3 mmol). The reaction wasstirred overnight at 40° C. After cooling to RT, the reaction wasquenched with water (20 mL) and ether (100 mL). The organic phase waswashed with water, brine, dried over magnesium sulfate and concentratedto afford the crude product which was purified by flash chromatographyon silica gel eluting with 0 to 10% MeOH in DCM. The solvent wasevaporated to dryness to afford2-(tert-butyldiphenylsilyloxy)-N-methylethanamine (4.4 g, 53%) as acolorless oil. NMR Spectrum (DMSOd6): 0.99 (s, 9H), 2.27 (s, 3H), 2.63(t, 2H), 3.68 (t, 2H), 7.40-7.49 (m, 6H), 7.60-7.65 (m, 4H).8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid was prepared as described in Example 2.00 and the two enantiomerswere separated by chiral preparative HPLC using the followingconditions:

Instrument Kronlab Column 100 mm Chiralpak IC 20 μm EluentDCM/IPA/HOAC/TEA 50/50/0.2/0.1 Oven Temperature Ambient Flow 350 ml/minWavelength 254 nm, 280 nm Sample Conc 4.5 g/100 ml in DCM/IPA 50/50Injection volume 100 mL Run Time 40 min

48.1 g of racemic compound was separated using the above conditions togive:

First eluting enantiomer 24 g (strength 76%) [α]^(D) _(20°): +115° inMeCN (enantiomer 1)

Second eluting enantiomer 24.1 g (strength 81%) [α]^(D) _(20°): −102° inMeCN (enantiomer 2)

1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (135 mg,0.71 mmol) was added in one portion to8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid (200 mg, 0.35 mmol, enantiomer 1; [α]^(D) _(20°): +115°),2-(tert-butyldiphenyl silyloxy)-N-methylethanamine (221 mg, 0.71 mmol)and 2-hydroxy-pyridine N-oxide (78 mg, 0.71 mmol) dissolved in DCM (2mL) under argon. The resulting solution was stirred at RT overnight. Thesolution was evaporated to dryness, water was added and product wasextracted with DCM. The organic phase was washed with brine and driedover magnesium sulfate, the solvent was evaporated to affordN-(2-(tert-butyldiphenylsilyloxy)ethyl)-8-(1-(3,5-difluorophenylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(94%) as a white solid. Mass Spectrum: M+H⁺ 726.

Lithium bis(trimethylsilyl)amide (0.539 mL, 0.54 mmol) was added to astirred solution ofN-(2-(tert-butyldiphenylsilyloxy)ethyl)-8-(1-(3,5-difluorophenylamino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(230 mg, 0.32 mmol) dissolved in dry THF (3 mL). The solution wasstirred over a period of 10 minutes at −20° C. under nitrogen. Dimethylsulfate (0.051 mL, 0.54 mmol) was added to the mixture and the resultingsuspension was left to rise to RT for 1 h30 under nitrogen. A sat. aq.sol. of NH₄Cl was added and the reaction mixture was extracted with DCM.The combined organic phases were washed with brine, dried over magnesiumsulfate and concentrated. The crude product was diluted with DCM andpurified by flash chromatography on silica gel eluting with 4% ethylalcohol in DCM. The solvent was evaporated to dryness to affordN-(2-(tert-butyldiphenylsilyloxy)ethyl)-8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamideenantiomer 1 (210 mg, 90%) as a white solid. Mass Spectrum: M+H⁺ 740.

EXAMPLE 7.0B8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(enantiomer 2)

This compound was prepared using an analogous procedure to thatdescribed for Example 7.0a but instead using enantiomer 2 of8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid ([α]^(D) _(20°): −102°). ThusN-(2-(tert-butyldiphenylsilyloxy)ethyl)-8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamideenantiomer 2 (1.8 g, 2.43 mmol) gave8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-N-(2-hydroxyethyl)-N-methyl-2-morpholino-4-oxo-4H-chromene-6-carboxamideenantiomer 2 (0.985 g, 80%) as a white solid [α]^(D) _(20°): +10° inMeCN. Mass Spectrum: M+H⁺ 502. NMR Spectrum (DMSOd6): 1.54 (bs, 3H),2.61 (s, 3H), 2.96 (s, 1.5H), 3.01 (s, 1.5H), 3.20-3.30 (m, 4H),3.41-3.58 (m, 7H), 3.65 (bs, 1H), 4.68 (bs, 1H), 5.54 (s, 1H), 5.58 (q,1H), 6.40 (d, 2H), 6.54 (t, 1H), 7.70 (bs, 0.5H), 7.77 (bs, 0.5H), 7.90(bs, 0.5H) 7.93 (bs, 0.5H).

EXAMPLE 7.01A8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(enantiomer 2)

Tetrabutylammonium fluoride (1N in THF) (7.83 mL, 7.83 mmol) was addeddropwise to a stirred solution of6-(4-(tert-butyldiphenylsilyloxy)piperidine-1-carbonyl)-8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4H-chromen-4-one(3 g, 3.92 mmol, enantiomer 2 in Example 7.0a, [α]^(D) _(20°): −102°)dissolved in THF (20 mL) at RT under nitrogen and stirred for 2 hrs. Themixture was evaporated to dryness, diluted with DCM, washed with water.The organic phase was washed with brine, dried over magnesium sulfate,filtered and concentrated. The crude product was purified by flashchromatography on silica gel (80 g) eluting with 3 to 7% MeOH in DCM.The solvent was evaporated to dryness to give a foam which wastriturated in ether (2-5 mL). The resulting white solid was collected byfiltration and dried to give8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-oneenantiomer 2 (1.7 g, 82%) as a white solid. Mass Spectrum: M+H⁺ 528.[α]^(D) _(20°): +7° in MeCN.

The preparation of the racemic8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-oneis described in Example 5.01.

The6-(4-(tert-butyldiphenylsilyloxy)piperidine-1-carbonyl)-8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-morpholino-4H-chromen-4-oneenantiomer 2 used as starting material was made using an analogousprocedure to the one described for the synthesis of the startingmaterial in Example 7.0a. Mass Spectrum: M+H⁺ 766.

EXAMPLE 7.01B8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-one(enantiomer 1)

This compound was prepared using an analogous procedure to thatdescribed in Example 7.01a except that the chiral acid starting materialwas8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid enantiomer 1 ([α]^(D) _(20°): +115°) from Example 7.0a. There wasthus obtained:8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-6-(4-hydroxypiperidine-1-carbonyl)-2-morpholino-4H-chromen-4-oneenantiomer 1 (45 mg, 69%) as a white solid. [α]^(D) _(20°): −3° in MeCN.Mass Spectrum: M+H⁺ 528.

EXAMPLE 7.026-(azetidine-1-carbonyl)-8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4H-chromen-4-one(enantiomer 2)

2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (409 mg, 1.36 mmol) was added portionwise to asuspension of the enantiomer 2 of8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4-oxo-4H-chromene-6-carboxylicacid ([α]^(D) _(20°): −102°, enantiomer 2, see Example 7.0a) (450 mg,1.05 mmol) and N-ethyl-N-isopropylpropan-2-amine (0.310 mL, 1.78 mmol)in DCM (5 mL) at RT under nitrogen. The resulting mixture was stirred atRT for 4 hrs. Azetidine (0.211 mL, 3.14 mmol) was then added to themixture and stirring was maintained overnight. The mixture was dilutedwith DMF and concentrated to remove DCM. The reaction mixture waspurified by preparative HPLC on a Waters X-Bridge system. The fractionswere evaporated to dryness to afford a white solid6-(azetidine-1-carbonyl)-8-(1-(3,5-difluorophenylamino)ethyl)-2-morpholino-4H-chromen-4-one(300 mg, 61%). Mass Spectrum: M+H⁺ 470. [α]^(D) _(20°): −113° in MeCN.NMR Spectrum (DMSOd6): 1.50 (d, 3H), 2.18-2.28 (m, 2H), 3.50-3.63 (m,4H), 3.69-3.78 (m, 4H), 3.98-4.07 (m, 2H), 4.09-4.19 (m, 2H), 4.97-5.05(m, 1H), 5.61 (s, 1H), 6.15 (d, 2H), 6.21 (t, 1H), 7.03 (d, 1H), 7.79(d, 1H), 8.02 (d, 1H).

EXAMPLE 8.08-(1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(single enantiomer)

To a mixture of8-(1-aminoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(350 mg, 1.01 mmol, single enantiomer, [α]^(D) _(20°): +35° inacetonitrile), cesium carbonate (1288 mg, 3.95 mmol),(9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine) (147 mg, 0.25mmol) and 1-bromo-3-chloro-5-fluorobenzene (467 mg, 2.23 mmol) indegassed 1,4-dioxane (2 ml), was addedtris(dibenzylideneacetone)dipalladium (70 mg, 0.08 mmol). The suspensionwas heated in a sealed container at 95° C. for 16 hrs. The reactionmixture was filtered through a short pad of dicalite and concentratedunder reduced pressure. The crude product was purified by flashchromatography on silica gel eluting with 0 to 8% isopropanol in DCM.The solvent was evaporated to dryness, the product triturated withdiethyl ether-DCM (9:1), collected by filtration and dried to afford8-(1-(3-chloro-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(320 mg; 67%) as a clear yellow solid. Mass Spectrum: M+H⁺ 474. [α]^(D)_(20°): −138°. NMR Spectrum (DMSOd6): 1.52 (d, 3H), 2.75 (bs, 3H), 2.95(bs, 3H), 3.49-3.63 (m, 4H), 3.70-3.79 (m, 4H), 4.98-5.07 (m, 1H), 5.60(s, 1H), 6.23 (d, 1H), 6.42 (d, 1H), 6.43 (ddd, 1H), 6.94 (d, 1H), 7.54(d, 1H), 7.80 (d, 1H).

The8-(1-aminoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(single enantiomer, [α]^(D) _(20°): +35° in acetonitrile) used asstarting material was made as follows:—

N-ethyl-N-isopropylpropan-2-amine (9.4 mL, 53.9 mmol) was added to8-acetyl-2-morpholino-4-oxo-4H-chromene-6-carboxylic acid (5.7 g, 18mmol) in DCM (100 mL). Five minutes later dimethylamine hydrochloride(2.9 g, 35.9 mmol) and2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (6.92 g, 21.6 mmol) were added. The mixture wasdiluted with DCM, washed with NaHCO3, dried over magnesium sulfate andconcentrated to afford the crude product which was purified by flashchromatography on silica gel eluting with 0 to 5% MeOH/DCM. The solventwas evaporated to dryness to afford8-acetyl-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide (2.75g, 46%) as a yellow solid.

Mass Spectrum: M+H⁺ 345.

Tetraethoxytitanium (12.5 g, 46.5 mmol) was added to a stirred solutionof 8-acetyl-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide (4g, 11.6 mmol) and (R)-2-methylpropane-2-sulfinamide (2.48 g, 20.4 mmol)in THF (100 mL) under nitrogen. The resulting mixture was stirred underreflux for 24 hrs. The reaction mixture was allowed to cool to RT,quenched with brine (100 ml) and diluted with ethyl acetate. Theprecipitate was removed by filtration over celite and washed with ethylacetate. The phases were separated and the aqueous phase was extractedwith ethyl acetate. The organic phases were combined, washed with water(twice), brine, dried over magnesium sulfate and concentrated to drynessto afford 2 g of the desired product. More product (2.8 g) was recoveredfrom the aqueous phase by DCM extraction. The 2 batches were combined togive8-(1-(tert-butylsulfinylimino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(4.5 g, 87%) as an yellow foam, which was used in the next step withoutfurther purification. Mass Spectrum: M+H⁺ 448.

The8-(1-(tert-butylsulfinylimino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidewas diluted in DCM (35 mL) and MeOH (35 mL), acetic acid (4.6 mL, 80.4mmol) and sodium cyanotrihydroborate (1.9 g, 30.2 mmol) was added at−15° C. The resulting mixture was stirred at −15° C. for 5 hrs thenallowed to warm to 0° C. A sat. sol. of Na2CO3 was added at 0° C. untilpH˜8-9 and extracted with DCM (×2). The organic phase was washed withbrine, dried over magnesium sulfate and concentrated. The crude productwas purified by flash chromatography on silica gel (column SiO2, 15-40μm-150 g Merck) eluting with 5 to 15% EtOH in DCM. The fractionscontaining impure product were concentrated down and repurified usingthe same system. The fractions containing pure8-(1-((R)-1,1-dimethylethylsulfinamido)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamidewere combined and the solvent was evaporated to dryness to afford 2.48g, (5.52 mmol, 54.9%) as a white foam. The fractions containing amixture of diastereoisomers were combined, concentrated and purified bypreparative HPLC on a Waters X-Bridge system. The fractions containingthe desired compound were evaporated to dryness to afford a further cropof8-(1-((R)-1,1-dimethylethylsulfinamido)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(0.25 g, 0.556 mmol, 5.53%) as a white foam. These 2 batches (2.48 g)and (0.25 g) were combined to give 8-(1-((R)-1,1-dimethyl ethylsulfinamido)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(2.73 g, 60%, diastereomeric excess>95%). Mass Spectrum: M+H⁺ 450.

Hydrogen chloride in dioxane 4M (15 mL, 60.1 mmol) was added to8-(1-((R)-1,1-dimethylethylsulfinamido)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(2.7 g, 6.0 mmol) dissolved in dioxane (40 mL). The resulting whitesuspension was stirred at RT for 1 hour, collected by filtration, washedwith diethyl ether and dried. It was dissolved into a 5% methanolicammonia (7 N) sol. in DCM (200 mL) and stirred for 5 minutes. Theprecipitate (NH4Cl) was removed by filtration and the filtrate wasconcentrated to dryness to afford8-(1-aminoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(single enantiomer, 1.85 g, 89%) as an off-white solid. Mass Spectrum:M+H⁺ 346. [α]^(D) _(20°): +35° in acetonitrile.

EXAMPLE 8.018-(1-(3-cyano-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(single enantiomer)

8-(1-aminoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(300 mg, 0.87 mmol, [α]^(D) _(20°): +35° in acetonitrile) was reactedwith 3-bromo-5-fluorobenzonitrile (382 mg, 1.91 mmol) using a proceduresimilar to the one described in Example 8.oo. Purification was done byflash chromatography on silica gel eluting with 0 to 5% MeOH in DCMfollowed by preparative HPLC on a Waters X-Bridge system. The fractionscontaining the desired compound were evaporated to dryness to afford8-(1-(3-cyano-5-fluorophenylamino)ethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(330 mg, 82%) as a clear white solid. Mass Spectrum: M+H⁺ 465. [α]^(D)_(20°): −128° in MeCN. NMR Spectrum (CDCl3): 1.51 (d, 3H), 2.91 (s, 3H),3.09 (s, 3H), 3.44-3.60 (m, 4H), 3.78-3.93 (m, 4H), 4.85-4.94 (m, 1H),5.17 (d, 1H), 5.56 (s, 1H), 6.38 (ddd, 1H), 6.51 (s, 1H), 6.61 (d, 1H),7.70 (d, 1H), 8.12 (d, 1H).

EXAMPLE 8.02N,N-dimethyl-2-morpholino-4-oxo-8-(1-(3,4,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamide(single enantiomer)

8-(1-aminoethyl)-N,N-dimethyl-2-morpholino-4-oxo-4H-chromene-6-carboxamide(232 mg, 0.67 mmol, [α]^(D) _(20°): +35° in acetonitrile),3,4,5-trifluorophenylboronic acid (236 mg, 1.34 mmol), diacetoxycopperhydrate (148 mg, 0.74 mmol) and molecular sieves 4A (1 g) were weighedout in a flask. Dichloroethane (4 mL) then pyridine (0.109 mL, 1.34mmol) were added and the resulting mixture was stirred at RT for 2 daysunder an oxygen atmosphere. The mixture was diluted with DCM, filteredthrough a pad of celite, the filtrate was washed with a 0.5N NaOH aq.sol. and the aqueous phase extracted with DCM. The combined organicswere dried over magnesium sulphate and concentrated down. The crudeproduct was purified by flash chromatography on silica gel eluting with0 to 10% MeOH in ethyl acetate/DCM (1:1). The solvent was evaporated todryness to affordN,N-dimethyl-2-morpholino-4-oxo-8-(1-(3,4,5-trifluorophenylamino)ethyl)-4H-chromene-6-carboxamidesingle enantiomer (100 mg, 31%) as a white foam. Mass Spectrum: M+H⁺476.[o]^(D) _(20°): −108° in MeCN. NMR Spectrum (CDCl3): 1.56 (d, 3H), 2.91(s, 3H), 3.09 (s, 3H), 3.47-3.56 (m, 4H), 3.82-3.90 (m, 4H), 4.35 (d,1H), 4.79-4.88 (m, 1H), 5.56 (s, 1H), 6.03 (dd, 2H), 7.70 (d, 1H), 8.12(d, 1H).

EXAMPLE 9.08-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide

8-(1-bromoethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamidehydrobromide (150 mg, 0.30 mmol) and 4-fluoroaniline (0.113 mL, 1.19mmol) in DMA (1 mL) were stirred at RT for 4 hrs. The reaction mixturewas filtered and purified by preparative HPLC using a reverse-phasecolumn (C-18, 5 microns silica, 19 mm diameter, 100 mm length, flow rateof 40 mL/minute) and decreasingly polar mixtures of water (containing0.2% ammonium carbonate) and acetonitrile as eluent. A furtherpurification was done by flash chromatography on silica gel eluting with0 to 10% MeOH in DCM. The solvent was evaporated to dryness to afford aoil which was triturated with pentane to give8-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(80 mg, 59%) as a white solid.

NMR Spectrum (DMSOd6): 1.16 (d, 3H), 1.50 (d, 3H), 2.66 (bs, 3H), 2.81(ddd, 1H), 2.93 (bs, 3H), 3.14 (ddd, 1H), 3.57-3.70 (m, 2H), 3.87-4.03(m, 3H), 4.93-5.01 (m, 1H), 5.62 (s, 1H), 6.24-6.29 (m, 1H), 6.43-6.50(m, 2H), 6.85 (t, 2H), 7.55 (d, 0.5H), 7.56 (d, 0.5H), 7.77 (d, 1H).

The8-(1-bromoethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamidehydrobromide used as starting material was made as follows:—

To a suspension of methyl 3-acetyl-5-bromo-4-hydroxybenzoate (75 g, 258mmol) in THF (350 mL) at −50° C. under nitrogen was added sodiumbis(trimethylsilyl)amide (1M in THF) (904 mL, 903.58 mmol) over a 15 minperiod. The dark solution was allowed to warm to −5-0° C. and stirredfor 1 h. Carbon disulfide (24.8 mL, 413 mmol) was added in one portionto the solution at −20° C. The mixture was allowed to warm to RT andstirred for 24 hrs. The reaction mixture was cooled to −50° C., quenchedslowly with a 15% aq. sol. H2SO4 (750 mL) (need to trap the H2S formed).The reaction was extracted 3 times with ethyl acetate. The organic phasewas washed with brine, dried over magnesium sulfate and concentrated.This residue was triturated with DCM (500 mL), collected by filtration,washed with ether and dried under vacuum to give methyl8-bromo-4-hydroxy-2-thioxo-2H-chromene-6-carboxylate (33.5 g, 41%) as anyellow solid. The filtrate was evaporated and the resulting dark gum wastriturated with ethyl acetate (300 mL) to give a solid which wascollected by filtration washed with ether and dried under vacuum to givea second batch of methyl8-bromo-4-hydroxy-2-thioxo-2H-chromene-6-carboxylate (17.5 g, 22%) as aorange solid. Mass Spectrum: [M−H]⁻ 314 for both batches.

Iodoethane (2.04 mL, 25.54 mmol), was added to a stirred suspension ofmethyl 8-bromo-4-hydroxy-2-thioxo-2H-chromene-6-carboxylate (2.3 g, 7.30mmol) and potassium carbonate (1.21 g, 8.76 mmol) in acetone (100 mL)under argon. The resulting mixture was stirred at 60° C. for 2 hrs. Themixture was concentrated in vacuo, and the residue was partioned betweenwater and DCM. The aqueous layer was extracted into DCM, and the organicextracts were combined, washed with brine, dried over magnesium sulfateand evaporated. The crude product was purified by flash chromatographyon silica gel eluting with 0 to 10% ethyl acetate in DCM. The solventwas evaporated to dryness to afford methyl8-bromo-2-(ethylthio)-4-oxo-4H-chromene-6-carboxylate (1.8 g, 72%) as anorange solid. Mass Spectrum: M+H⁺ 343.

To a stirred solution of methyl8-bromo-2-(ethylthio)-4-oxo-4H-chromene-6-carboxylate (1.8 g, 5.24 mmol)in DCM (40 mL) was added dropwise 3-chlorobenzoperoxoic acid (2.59 g,10.49 mmol) while keeping the temperature around 20° C. with a cold baththen left to stir at RT for 2 hrs. The solution was cooled to −15° C.,the solid was filtered off and rinsed with cold DCM. The filtrate wasthen washed with a solution of sodium sulfothioate pentahydrate (0.651g, 2.62 mmol) in 30 mL H2O, and twice with a solution of NaHCO3. Theorganic was dried over magnesium sulfate and evaporated to afford methyl8-bromo-2-(ethylsulfonyl)-4-oxo-4H-chromene-6-carboxylate (1.85 g, 94%)as a red powder consisting approximately of a 70:30 sulfone/sulfoxidemixture which was used as such for the next step.

A mixture of (R)-2-methylmorpholine hydrochloride (0.436 g, 3.17 mmol)and N-ethyl-N-isopropylpropan-2-amine (1.184 mL, 6.80 mmol) in DCM (5mL) was added dropwise to a stirred solution of methyl 8-bromo-2-(ethylsulfonyl)-4-oxo-4H-chromene-6-carboxylate (0.85 g, 2.27 mmol) in DCM (10mL) at 10° C. under argon. The resulting solution was stirred at RT for3 hrs. The reaction mixture was quenched with 1M HCl, the phases wereseparated, the organic phase was washed with brine dried over magnesiumsulfate and concentrated to afford the crude product which was purifiedby flash chromatography on silica gel eluting with 0 to 10% MeOH in DCM.The solvent was evaporated to dryness to afford after a trituration withether, methyl8-bromo-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate(0.760 g, 88%) as a pale yellow foam. Mass Spectrum: M+H⁺ 382. Thereaction was repeated on a similar scale before proceeding to the nextstep.

Methyl8-bromo-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate (1.45g, 3.79 mmol), bis(triphenylphosphine) palladium(II) chloride (0.107 g,0.15 mmol) and tributyl(1-ethoxyvinyl)stannane (1.346 mL, 3.98 mmol) in1,4-dioxane (20 mL) were degassed, purged with argon and heated to 90°C. for 4 hrs. After cooling to RT, HCl 2N (1.9 mL, 3.79 mmol) was addedand the mixture was left to stir for 1 h. The reaction mixture wasconcentrated, suspended in water, neutralised with NaHCO3 and extractedwith DCM. The combined organic phases were washed with water, brine,dried over magnesium sulfate and concentrated. The crude product wastriturated with n-heptane, filtered and retriturated with ether,filtered and dried to give methyl8-acetyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate(1.15 g, 88%) as a grey solid. Mass Spectrum: M+H⁺ 346.

To a solution of methyl8-acetyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate(1.15 g, 3.33 mmol) in MeOH (20 mL)/DCM (10 mL) was added sodiumtetrahydroborate (0.139 g, 3.66 mmol) at −10° C. The reaction mixturewas quenched with water (50 mL) 15 min later. The volatiles were removedand the aqueous layer was extracted twice with DCM. Combined organicsphases were washed with brine, dried over magnesium sulfate andconcentrated. The residue was triturated with MTBE/DCM (9/1) andcollected by filtration to give methyl8-(1-hydroxyethyl)-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate(1.0 g, 86%) as a grey solid, which was used in the next step withoutfurther purification. Mass Spectrum: M+H⁺ 348.

NaOH (3.56 mL, 7.11 mmol) was added to a stirred suspension of methyl8-(1-hydroxyethyl)-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate(988 mg, 2.84 mmol) in MeOH (10 mL)/water (10 mL). The resulting mixturewas stirred at RT for 1 hour then acidified to pH 2-3 with a 2N aq. HCl(7.68 mL, 7.68 mmol). The resulting precipitate was collected byfiltration, washed with diethyl ether and dried to afford8-(1-hydroxyethyl)-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylicacid (900 mg, 95%) as a grey solid, which was used without furtherpurification. Mass Spectrum: M+H⁺ 334.

2-(2,5-dioxopyrrolidin-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (2.38 g, 3.96 mmol) was added portionwise to8-(1-hydroxyethyl)-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylicacid (0.88 g, 2.64 mmol) and N-ethyl-N-isopropylpropan-2-amine (1.380mL, 7.92 mmol) suspended in DCM (15 mL) at 10° C. under nitrogen. Theresulting mixture was stirred at RT for 2 hrs. Dimethylamine (3.96 mL,7.92 mmol) was then added at 10° C. and the resulting mixture wasstirred at RT overnight. The mixture was poured onto a silica gel columnand purified by flash chromatography eluting with 0 to 10% methanolicammonia (7 N) in DCM. The solvent was evaporated to dryness to afford afoam which crystallised from ethyl acetate, the solid was collected byfiltration and dried to a constant weight in a vacuum oven to afford8-(1-hydroxyethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(0.800 g, 84%) as a white solid. Mass Spectrum: M+H⁺ 361.

A solution of tribromophosphine (0.215 mL, 2.29 mmol) in1,2-dichloroethane (1 mL) at 10° C. was added dropwise to8-(1-hydroxyethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(750 mg, 2.08 mmol) suspended in 1,2-dichloroethane (9 mL) undernitrogen. The resulting suspension was stirred at 50° C. for 1 hour. Thereaction mixture was allowed to cool to RT under stirring and dilutedwith diethyl ether (40 mL). The precipitate was collected by filtration,washed with diethyl ether and dried to a constant weight to afford8-(1-bromoethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamidehydrobromide (1.3 g, 124%) as a white solid, which was used withoutfurther purification. Mass Spectrum: M+H⁺ 424.

EXAMPLE 9.018-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide

8-(1-bromoethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamidehydrobromide (1 g, 1.98 mmol) and 3,5-difluoroaniline (1.024 g, 7.93mmol) in DMA (5 mL) were reacted as described in Example 9.0 to give8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(0.550 g, 57%). Mass Spectrum: M+H⁺ 472. NMR Spectrum (DMSOd6): 1.16 (d,3H), 1.52 (d, 3H), 2.74 (bs, 3H), 2.75-2.84 (m, 1H), 2.95 (bs, 3H),3.08-3.16 (m, 1H), 3.56-3.68 (m, 2H), 3.86-4.04 (m, 3H), 4.97-5.08 (m,1H), 5.62 (s, 1H), 6.12-6.19 (m, 2H), 6.22 (t, 1H), 6.93 (d, 0.5H), 6.94(d, 0.5H), 6.54 (d, 0.5H), 6.55 (d, 0.5H), 7.81 (s, 1H).

The above mixture of diastereoisomers (495 mg) was purified by chiralpreparative HPLC using the following conditions:

Column CelluCoat 250 × 50 10 μm Eluent Heptane/IPA/TEA 50/50/0.1 OvenTemperature Ambient Flow 120 mL/min Wavelength 270 nm Sample Conc 50mg/ml EtOH/DCM 1:1 Injection amount 495 mgFirst eluting diastereoisomer 234 mg (Example 9.01a) [α]^(D) _(20°):+136° in MeCN.Second eluting diastereoisomer 240 mg (Example 9.01b) [α]^(D) _(20°):−99° in MeCN.

EXAMPLE 9.028-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide

A solution of8-(1-bromoethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(670 mg, 1.42 mmol) and 3,5-difluoroaniline (736 mg, 5.70 mmol) in DMA(4 mL) were stirred at 50° C. for 4 hrs then at rt over the weekend. Thereaction mixture was diluted with ethyl acetate, washed with a sat. aq.sol. of sodium hydrogenocarbonate, water brine, dried over magnesiumsulfate and concentrated to afford the crude product, which was purifiedby flash chromatography on silica gel eluting with 0 to 10% MeOH in DCM.The solvent was evaporated to dryness and the obtained foam wastriturated with diethyl ether to give a white solid which was collectedby filtration and dried under vacuum to give8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(400 mg, 60%). Mass Spectrum: M+H⁺ 472. NMR Spectrum (CDCl3): 1.25 (d,3H), 1.61 (d partially hidden by H2O, 3H), 2.83 (ddd, 1H), 2.90 (bs,3H), 3.08 (bs, 3H), 3.18 (dddd, 1H), 3.63-3.82 (m, 4H), 4.02 (ddd, 1H),4.37 (bs, 1H), 4.88-4.97 (m, 1H), 5.55 (s, 1H), 5.97 (d, 2H), 6.13 (t,1H), 7.72 (d, 1H), 8.12 (d, 1H).

The above mixture of diastereoisomers (370 mg) was purified by chiralpreparative HPLC using the following conditions:

Column Chiralpak IC 20 × 250 mm, 10 μm Eluent DCM/IPA 6:4 OvenTemperature Ambient Flow 20 mL/min Wavelength 280 nm Sample Conc 110mg/mL in DCM/MeOH 6:4 Injection 55 mgFirst eluting diastereomer 122 mg (Example 9.02a) [α]^(D) _(20°): +111°in MeCNSecond eluting diastereomer 105 mg (Example 9.02b) [α]^(D) _(20°): −163°in MeCN

EXAMPLE 9.038-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide

8-(1-bromoethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(110 mg, 0.20 mmol) and 4-fluoroaniline in DMA (1 mL) were stirred at50° C. for 5 hrs. The reaction mixture was purified by preparative HPLCon a Waters X-Bridge system. The fractions containing the desiredcompound were concentrated. The obtained gum was triturated in diethylether and petroleum ether, the resulting solid was collected byfiltration and dried to afford8-(1-(4-fluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(45 mg, 51%) as a white solid. Mass Spectrum: M+H⁺ 545. NMR Spectrum(CDCl3): 1.25 (d, 1.5H), 1.26 (d, 1.5H), 1.61 (d, 3H), 2.77-2.88 (m,1H), 2.83 (bs, 3H), 3.06 (bs, 3H), 3.13-3.22 (m, 1H), 3.64-3.82 (m, 4H),3.97 (bs, 1H), 3.98-4.06 (m, 1H), 4.91 (q, 1H), 5.56 (s, 1H), 6.37-6.43(m, 2H), 6.82 (t, 2H), 7.73 (d, 1H), 8.11 (d, 1H).

EXAMPLE 10.0

For preparation of the compounds of Examples 10.01 to 10.03 (shown inTable III), TBTU (96 mg, 0.30 mmol) was added in one portion to astirred solution of the desired amine reagent (-, 0.30 mmol),8-(1-((3,5-difluorophenyl)(methyl)amino)ethyl)-2-((R)-2-methylmorpholino)-4-oxo-4H-chromene-6-carboxylicacid (125 mg, 0.27 mmol) and 4-methylmorpholine (0.066 mL, 0.60 mmol) inDMF (1 mL). The resulting solution was stirred at RT overnight. Thereaction mixture was filtered and purified by preparative HPLC on aWaters X-Bridge system. The fractions containing the desired compoundwere evaporated to dryness.

TABLE III Amine Prod. Ex. reageant Structure Product Mass Yield MH⁺10.01 dimethyl- amine

8-(1-((3,5- difluorophenyl)(methyl) amino)ethyl)-N,N- dimethyl-2-((R)-2-methylmorpholino)-4- oxo-4H-chromene-6- carboxamide 83 mg 63% 486 10.022- (methyl- amino) ethanol

8-(1-((3,5- difluorophenyl)(methyl) amino)ethyl)-N-(2- hydroxyethyl)-N-methyl-2-((R)-2- methylmorpholino)-4- oxo-4H-chromene-6- carboxamide 79mg 56% 516 10.03 piperidin- 4-ol

8-(1-((3,5- difluorophenyl)(methyl) amino)ethyl)-6-(4-hydroxypiperidine-1- carbonyl)-2-((R)-2- methylmorpholino)-4H-chromen-4-one 85 mg 58% 542Notes Further characterising data for the products is given below.

EXAMPLE 10.01

DMSOd6 at 323° K: 1.01 (d, 1.5H), 1.02 (d, 1.5H), 1.57 (d, 3H), 2.63 (s,1.5H), 2.67 (s, 1.5H), 2.65-2.71 (m, 0.5H), 2.74-2.83 (m, 0.5H), 2.98(bs, 6H), 3.31-3.55 (m, 2H), 3.61-3.81 (m, 4H), 5.56 (s, 1H), 5.59 (q,1H), 6.37 (t, 1H), 6.57 (d, 2H), 7.70 (s, 1H), 7.92 (s, 1H).

EXAMPLE 10.02

DMSOd6 at 323° K: 1.01 (d, 1.5H), 1.02 (d, 1.5H), 1.56 (d, 3H),2.46-2.51 (m, 0.5H), 2.61 (s, 1.5H), 2.64 (s, 1.5H), 2.64-2.71 (m,0.5H), 2.74-2.82 (m, 0.5H), 2.94-3.00 (m, 0.5H), 3.00 (bs, 3H),3.27-3.90 (m, 9H), 4.74 (t, 1H), 5.55 (s, 1H), 5.59 (q, 1H), 6.37 (t,1H), 6.55 (d, 2H), 7.74 (bs, 1H), 7.93 (s, 1H).

EXAMPLE 10.03

DMSOd6 at 323° K: 1.02 (d, 1.5H), 1.05 (d, 1.5H), 1.39 (bs, 2H), 1.58(d, 3H), 1.79 (bs, 2H), 2.48-2.55 m partially hidden by DMSOd6, 0.5H),2.64 (s, 1.5H), 2.66-2.71 (m, 0.5H), 2.67 (s, 1.5H), 2.76-2.84 (m,0.5H), 2.95-3.03 (m, 0.5H), 3.21 (bs partially hidden by H2O, 2H),3.31-3.56 (m, 2H), 3.63-3.81 (m, 4H), 3.84 (bs, 2H), 4.69 (d, 1H), 5.56(s, 1H), 5.58 (q, 1H), 6.37 (t, 1H), 6.53 (d, 2H), 7.64 (bs, 1H), 7.89(s, 1H).

EXAMPLE 118-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide

2-(1H-benzo[d][1,2,3]triazol-1-yl)-1,1,3,3-tetramethylisouroniumtetrafluoroborate (55.6 mg, 0.17 mmol), was added in one portion to astirred solution of8-(1-(3,5-difluorophenylamino)ethyl)-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxylicacid (70 mg, 0.16 mmol), dimethylamine (0.095 mL, 0.19 mmol) and4-methylmorpholine (0.038 mL, 0.35 mmol) in DMF (1 mL). The resultingsolution was stirred at RT for 1 hour. The reaction mixture was purifiedby preparative HPLC using a reverse-phase column (C-18, 5 micronssilica, 19 mm diameter, 100 mm length, flow rate of 40 mL/minute) anddecreasingly polar mixtures of water (containing 0.2% ammoniumcarbonate) and acetonitrile as eluent. The fractions containing thedesired compound were evaporated to dryness to afford a residue whichwas triturated with Et2O, filtered and dried to give8-(1-(3,5-difluorophenylamino)ethyl)-N,N-dimethyl-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxamide(45 mg, 61%) as a pale orange solid. Mass Spectrum: M+H⁺ 471. NMRSpectrum (CDCl3): 1.37 (d, 1.5H), 1.41 (d, 1.5H), 1.59 (d, 1.5H), 1.60(d, 1.5H), 2.89 (s, 1.5H), 2.91 (s, 1.5H), 3.08 (s, 3H), 3.35-3.43 (m,1H), 3.52-3.69 (m, 2H), 3.75-3.83 (m, 2H), 3.98-4.09 (m, 2H), 4.52 (d,1H), 4.85-4.96 (m, 1H), 5.53 (s, 1H), 5.97 (d, 2H), 6.11 (t, 1H), 7.71(d, 0.5H), 7.73 (d, 0.5H), 8.13 (d, 1H).

The8-(1-(3,5-difluorophenylamino)ethyl)-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxylicacid used as starting material was made as follows:—

Methyl 8-bromo-2-(ethylthio)-4-oxo-4H-chromene-6-carboxylate (850 mg,2.29 mmol, prepared as described in Example 9) and 3,5-difluoroaniline(621 mg, 4.81 mmol) in DMA (10 mL) were stirred at 50° C. overnight. Thereaction mixture was diluted with water/ethyl acetate. The organic layerwas washed with brine, dried over MgSO4 and concentrated. The crudeproduct was purified by flash chromatography on silica gel eluting with0 to 25% ethyl acetate in dichloromethane. The solvent was evaporated todryness to afford methyl8-(1-(3,5-difluorophenylamino)ethyl)-2-(ethylthio)-4-oxo-4H-chromene-6-carboxylate(550 mg, 57%) as a off-white solid. Mass Spectrum: M+H⁺ 420.

3-chlorobenzoperoxoic acid (326 mg, 1.13 mmol) was added in one portionto a stirred suspension of methyl8-(1-(3,5-difluorophenylamino)ethyl)-2-(ethylthio)-4-oxo-4H-chromene-6-carboxylate(475 mg, 1.13 mmol) in DCM (5 mL) cooled with an water/ice bath. Theresulting mixture was stirred at RT for 1 h. The suspension was cooledto −15° C. and filtered, the solid was washed with cold DCM (5 mL). Thefiltrate was then washed with an aq. sol. of sodium thiosulfatepentahydrate in water (10 mL), and with a mixture of a sat. sol. ofNaHCO3 and water (1:1, 15 mL). The organic layer was decanted, driedover MgSO4 and evaporated to afford the crude product methyl8-(1-(3,5-difluorophenylamino)ethyl)-2-(ethylsulfinyl)-4-oxo-4H-chromene-6-carboxylate(500 mg, 100%) as a reddish foam. Mass Spectrum: M+H⁺ 436.

(S)-3-methylmorpholine (87 mg, 0.86 mmol) was added to a stirredsolution of methyl8-(1-(3,5-difluorophenylamino)ethyl)-2-(ethylsulfinyl)-4-oxo-4H-chromene-6-carboxylate(250 mg, 0.57 mmol) and N-ethyl-N-isopropylpropan-2-amine (0.150 mL,0.86 mmol) in acetonitrile (3 mL) at RT. The resulting brown mixture wasstirred at 75° C. for 7 hours. The reaction mixture was concentrated,diluted with DCM, washed with a 1M hydrochloric acid 1M, brine driedover magnesium sulfate and concentrated to afford the crude productwhich was purified by flash chromatography on silica gel eluting with 0to 5% MeOH in DCM. The solvent was evaporated to dryness to affordmethyl8-(1-(3,5-difluorophenylamino)ethyl)-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate(90 mg, 34%) as a yellow foam. Spectrum: [M−H]⁻ 457.

2N NaOH (0.206 mL, 0.41 mmol) was added to a stirred suspension ofmethyl8-(1-(3,5-difluorophenylamino)ethyl)-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxylate(90 mg, 0.20 mmol) in MeOH (1 mL)/THF (1 mL) and cooled with anice-water bath. The resulting suspension was stirred at RT for 2 hours.The reaction was incomplete thus the temperature was increased to 35° C.and stirred for an additional hour. The reaction mixture was cooled withan ice bath, the pH was adjusted to 2-3 with hydrochloric acid 1M andthe volatils were evaporated. The resulting precipitate was collected byfiltration, dried, triturated with diethyl ether, collected byfiltration and dried again under vacuum to give8-(1-(3,5-difluorophenylamino)ethyl)-2-((S)-3-methylmorpholino)-4-oxo-4H-chromene-6-carboxylicacid (75 mg, 86%). Mass Spectrum: M+H⁺ 445.

The invention claimed is:
 1. A compound of the formula XVI:

in which: R¹ is H or (1-4C)alkyl optionally substituted by 1, 2 or 3substituents independently selected from halogeno, hydroxy or(1-3C)alkoxy; R² is (1-4C)alkyl or (1-4C)alkoxy, either of which can beoptionally substituted by 1, 2 or 3 substituents independently selectedfrom halogeno, hydroxy, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy,cyano, (1-3C)alkylamino or di-[(1-3C)alkyl]amino; or R¹ and R² togetherform a 3 to 8 membered nitrogen containing heterocyclyl ring system,which optionally contains 1 or 2 further heteroatoms selected fromoxygen, nitrogen and sulphur, wherein a ring sulphur atom is optionallyoxidised to form the S-oxide(s), said ring being optionally substitutedby 1, 2 or 3 substituents independently selected from halogeno, hydroxy,(1-3C)alkyl, (2-3C)alkenyl, (2-3C)alkynyl, (1-3C)alkoxy, oxo,hydroxy-(1-3C)alkyl, halogeno-(1-3C)alkyl and (1-3C)alkoxy-(1-3C)alkyl;n is 0, 1, 2, 3 or 4; and each R⁹ group is (1-3C)alkyl.
 2. A compound ofthe Formula XVI as claimed in claim 1 in which R¹ is methyl; R² ismethyl; and n is 0.